Review



tuberin tsc2 d93f12 xp rabbit mab  (Cell Signaling Technology Inc)


Bioz Verified Symbol Cell Signaling Technology Inc is a verified supplier
Bioz Manufacturer Symbol Cell Signaling Technology Inc manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
    • 86
      Buy from Supplier

    Structured Review

    Cell Signaling Technology Inc tuberin tsc2 d93f12 xp rabbit mab
    Tuberin Tsc2 D93f12 Xp Rabbit Mab, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/tsc2+d93f12/pmc13068069-4-0-6
    Average 86 stars, based on 1 article reviews
    tuberin tsc2 d93f12 xp rabbit mab - by Bioz Stars, 2026-09
    86/100 stars

    Images

    Related Articles

    Activation Assay:

    Article Title: Trans‐gnetin H isolated from the seeds of Paeonia species induces autophagy via inhibiting mTORC1 signalling through AMPK activation
    Article Snippet: The WB protocol was the same as our previous work.The WB protocol was the same as our previous work.. The primary antibodies Actin (20536‐1‐AP), LC3II (18725‐1‐AP), p62 (18420‐1‐AP) were purchased from Proteintech; TFEB (37785 S), pT389‐S6K (9234 S), p‐S6 (4858 S), S6K (9202 S), S6 (2217 S), p‐AMPKα (50081 S), AMPKα (5832 S), Raptor (2280 S) and TSC2 (D93F12) were purchased from Cell Signaling Technology; Flag (db7002) and HA (db2603) were purchased from Hangzhou Bio Technolog.. The secondary antibodies were obtained from Sigma.The secondary antibodies were obtained from Sigma.

    Article Title: A translational program that suppresses metabolism to shield the genome
    Article Snippet: Blots were stripped and reprobed when looking at total versus modified proteins unless otherwise noted.. Antibodies (all 1:1000): GLUT1 (Novus Biologicals, # NB110-39113), PAI-1 (Proteintech, #13801-1-AP), Puromycin (3RH11) (Kerafast, #EQ0001), β-ACTIN (C4) (SantaCruz Biotechnology, #sc-47778), EIF5A (abcam, #ab32443), EIF3D (Proteintech, # 10219-1-AP), EIF3K (Proteintech, # 10640-1-AP), acetyl-EIF5A (Lys47) (Boster Bio, #P01727), CYR61 (Proteintech, #26689-1-AP), SDC4 (R&D Systems, #P31431), SIRT1 (Proteintech, #13161-1-AP), SIRT2 (Proteintech, #19655-1-AP), NFKB2 (Proteintech, #10409-2-AP), 4E-BP (Cell Signaling, #9452 S), Phospho-4E-BP (Ser65) (Cell Signaling, #9451 S), TSC2 (D93F12) (Cell Signaling, #4308S), gamma H2AX (Novus, NB100-384), H2AX (Abcam, Ab11175), CRM-1 (Novus, NB100-79802), XPO-4 (Abcam, ab133237), eIF5A2 (Thermo, #PA5-30770), Phospho-RAPTOR (Ser792) (Cell Signaling, #89146S), RAPTOR (Cell Signaling, #2280), Phospho-Tuberin/TSC2 (Ser1387) (Cell Signaling, #5584S), NDRG1 (Abcam, #ab37897), and Anti-hypusine (mAbHpu24) antibody was kindly provided by Genentech.. The eIF5A1 antibodies and siRNAs used in this study were specific to eIF5A1 and not eIF5A2 as described by the manufacturer and validated in Supplementary Fig. .

    Article Title: Identification of mechanically regulated phosphorylation sites on tuberin (TSC2) that control mechanistic target of rapamycin (mTOR) signaling
    Article Snippet: Images were quantified using ImageJ software (U.S. National Institutes of Health, Bethesda, MD; http://rsb.info.nih.gov/nih-image/).Images were quantified using ImageJ software (U.S. National Institutes of Health, Bethesda, MD; http://rsb.info.nih.gov/nih-image/).. Antibodies - P-p70(389) (1A5) (#9206, used to probe Myc-tagged P-p70), P-p70(389) (#9234, used to probe endogenous P-p70), total p70 (49D7) (#2708), Anti-P-Akt substrate (RxRxxS*/T*) (23C8D2) for detection of PTSC2 (#10001), P-JNK(Thr183/Tyr185) (98F2) (#4671), total JNK (#9252), total mTOR (#2972), total Rictor (D16H9) (#9476), P-PKB (Ser473) (#4060), P-PKB (Thr308) (#9275S), total PKB (#9272), P-S6 (Ser240/4) (#5364), P-S6 (Ser235/6) (#2211), total S6 (#2217S), total TSC1 (D43E2) (#6935), total TSC2 (D93F12) (#4308) were purchased from Cell Signaling Technology (Danvers, MA, USA).. Anti-puromycin (12D10) (#MABE343) was purchased from EMD Millipore (Billerica, MA, USA).Anti-puromycin (12D10) (#MABE343) was purchased from EMD Millipore (Billerica, MA, USA).

    Article Title: Chronic signaling via the metabolic checkpoint kinase mTORC1 induces macrophage granuloma formation and marks sarcoidosis progression
    Article Snippet: HRP-conjugated secondary antibodies (Bethyl) 1:10000 were applied for 1 hour at room temperature in 4% low-fat milk and proteins were visualized using ECL substrate (Thermo Scientific).HRP-conjugated secondary antibodies (Bethyl) 1:10000 were applied for 1 hour at room temperature in 4% low-fat milk and proteins were visualized using ECL substrate (Thermo Scientific).. Primary antibodies were TSC2 (D93F12) 1:1000, p-Akt S473 (D9E) 1:1000, 4E-BP1 (53H11) 1:1000, p-S6 S240/244 (rabbit polyclonal) 1:3000, S6 (54D2) 1:1000, p-Akt T308 (D25E6) 1:1000, pan-Akt (40D4) 1:1000, β-tubulin (9F3) 1:1000 , p-RB Ser807/811 (D20B12) 1:1000, PDCD4 (D29C6) 1:1000, cleaved caspase 3 Asp175 (5A1E or rabbit polyclonal) 1:500, survivin (71G4B7) 1:1000 and p-S6K1 T389 (108D2) 1:1000, p-TSC2 S939 (rabbit polyclonal) 1:1000 obtained from Cell Signaling Technology.. Bcl-2 (N-19) 1:500, cyclin D1 (M-20) 1:500, p27Kip1 (C-19) 1:1000, cyclin A (C-19) 1:500, CDK4 (C-22) 1:1000, CDK6 (H-96) 1:500, and E2F1 (KH95) 1:500 were fom Santa Cruz. α-tubulin (CP06) 1:3000 was purchased from Calbiochem and GAPDH (rabbit polyclonal) 1:10000 from Trevigen.Bcl-2 (N-19) 1:500, cyclin D1 (M-20) 1:500, p27Kip1 (C-19) 1:1000, cyclin A (C-19) 1:500, CDK4 (C-22) 1:1000, CDK6 (H-96) 1:500, and E2F1 (KH95) 1:500 were fom Santa Cruz. α-tubulin (CP06) 1:3000 was purchased..

    Article Title: TSC1 promotes i NKT cell anergy and inhibits i NKT cell-mediated anti-tumor immunity
    Article Snippet: Fluorescence-conjugated anti-mouse TCRβ (H57-597), NK1.1 (PK136), CD44(IM7), B220(RA3-6B2), Gr-1(RB6-8C5), CD11c(N418), CD11b(M1/70), CD4 (GK1.5), CD8 (53-6.7), IFN-γ (XMG1.2), IL-4 (11B11), PD-1(RMP1-30), and ICOS(D10.G4.1) antibodies were purchased from BioLegend.Fluorescence-conjugated anti-mouse TCRβ (H57-597), NK1.1 (PK136), CD44(IM7), B220(RA3-6B2), Gr-1(RB6-8C5), CD11c(N418), CD11b(M1/70), CD4 (GK1.5), CD8 (53-6.7), IFN-γ (XMG1.2), IL-4 (11B11), PD-1(RMP1-30), and ICOS(D10.G4.1) antibodies were purchased from BioLegend.. Anti–phospho-p44/42 mitogen-activated protein kinase T202/Y204 (197G2), anti–phospho-S6 S240/244 (D68F8), anti–phospho-4E-BP1 T37/46 (236B4), anti–phospho-Akt S473 (193H12), TSC1 (D43E2), and TSC2 (D93F12) were purchased from Cell Signaling Technology, Inc. Anti–β-actin antibody was purchased from Sigma-Aldrich.. Cell surface staining was performed with 2% FBS-PBS.Cell surface staining was performed with 2% FBS-PBS.

    Article Title: TSC1 promotes i NKT cell anergy and inhibits i NKT cell-mediated anti-tumor immunity
    Article Snippet: CD11b(M1/70), CD4 (GK1.5), CD8 (53-6.7), IFN-γ (XMG1.2), IL-4 (11B11), PD-1(RMP1-30), and ICOS(D10.G4.1) antibodies were purchased from BioLegend. .. Anti–phospho-p44/42 mitogen-activated protein kinase T202/Y204 (197G2), anti–phospho-S6 S240/244 (D68F8), anti–phospho-4E-BP1 T37/46 (236B4), anti–phospho-Akt S473 (193H12), TSC1 (D43E2), and TSC2 (D93F12) were purchased from Cell Signaling Technology, Inc. Anti–β-actin antibody was purchased from Sigma-Aldrich. .. Cell surface staining was performed with 2% FBS-PBS.

    Transfection:

    Article Title: Trans‐gnetin H isolated from the seeds of Paeonia species induces autophagy via inhibiting mTORC1 signalling through AMPK activation
    Article Snippet: The WB protocol was the same as our previous work.The WB protocol was the same as our previous work.. The primary antibodies Actin (20536‐1‐AP), LC3II (18725‐1‐AP), p62 (18420‐1‐AP) were purchased from Proteintech; TFEB (37785 S), pT389‐S6K (9234 S), p‐S6 (4858 S), S6K (9202 S), S6 (2217 S), p‐AMPKα (50081 S), AMPKα (5832 S), Raptor (2280 S) and TSC2 (D93F12) were purchased from Cell Signaling Technology; Flag (db7002) and HA (db2603) were purchased from Hangzhou Bio Technolog.. The secondary antibodies were obtained from Sigma.The secondary antibodies were obtained from Sigma.

    Article Title: A translational program that suppresses metabolism to shield the genome
    Article Snippet: Blots were stripped and reprobed when looking at total versus modified proteins unless otherwise noted.. Antibodies (all 1:1000): GLUT1 (Novus Biologicals, # NB110-39113), PAI-1 (Proteintech, #13801-1-AP), Puromycin (3RH11) (Kerafast, #EQ0001), β-ACTIN (C4) (SantaCruz Biotechnology, #sc-47778), EIF5A (abcam, #ab32443), EIF3D (Proteintech, # 10219-1-AP), EIF3K (Proteintech, # 10640-1-AP), acetyl-EIF5A (Lys47) (Boster Bio, #P01727), CYR61 (Proteintech, #26689-1-AP), SDC4 (R&D Systems, #P31431), SIRT1 (Proteintech, #13161-1-AP), SIRT2 (Proteintech, #19655-1-AP), NFKB2 (Proteintech, #10409-2-AP), 4E-BP (Cell Signaling, #9452 S), Phospho-4E-BP (Ser65) (Cell Signaling, #9451 S), TSC2 (D93F12) (Cell Signaling, #4308S), gamma H2AX (Novus, NB100-384), H2AX (Abcam, Ab11175), CRM-1 (Novus, NB100-79802), XPO-4 (Abcam, ab133237), eIF5A2 (Thermo, #PA5-30770), Phospho-RAPTOR (Ser792) (Cell Signaling, #89146S), RAPTOR (Cell Signaling, #2280), Phospho-Tuberin/TSC2 (Ser1387) (Cell Signaling, #5584S), NDRG1 (Abcam, #ab37897), and Anti-hypusine (mAbHpu24) antibody was kindly provided by Genentech.. The eIF5A1 antibodies and siRNAs used in this study were specific to eIF5A1 and not eIF5A2 as described by the manufacturer and validated in Supplementary Fig. .

    Article Title: Identification of mechanically regulated phosphorylation sites on tuberin (TSC2) that control mechanistic target of rapamycin (mTOR) signaling
    Article Snippet: Images were quantified using ImageJ software (U.S. National Institutes of Health, Bethesda, MD; http://rsb.info.nih.gov/nih-image/).Images were quantified using ImageJ software (U.S. National Institutes of Health, Bethesda, MD; http://rsb.info.nih.gov/nih-image/).. Antibodies - P-p70(389) (1A5) (#9206, used to probe Myc-tagged P-p70), P-p70(389) (#9234, used to probe endogenous P-p70), total p70 (49D7) (#2708), Anti-P-Akt substrate (RxRxxS*/T*) (23C8D2) for detection of PTSC2 (#10001), P-JNK(Thr183/Tyr185) (98F2) (#4671), total JNK (#9252), total mTOR (#2972), total Rictor (D16H9) (#9476), P-PKB (Ser473) (#4060), P-PKB (Thr308) (#9275S), total PKB (#9272), P-S6 (Ser240/4) (#5364), P-S6 (Ser235/6) (#2211), total S6 (#2217S), total TSC1 (D43E2) (#6935), total TSC2 (D93F12) (#4308) were purchased from Cell Signaling Technology (Danvers, MA, USA).. Anti-puromycin (12D10) (#MABE343) was purchased from EMD Millipore (Billerica, MA, USA).Anti-puromycin (12D10) (#MABE343) was purchased from EMD Millipore (Billerica, MA, USA).

    Article Title: Chronic signaling via the metabolic checkpoint kinase mTORC1 induces macrophage granuloma formation and marks sarcoidosis progression
    Article Snippet: HRP-conjugated secondary antibodies (Bethyl) 1:10000 were applied for 1 hour at room temperature in 4% low-fat milk and proteins were visualized using ECL substrate (Thermo Scientific).HRP-conjugated secondary antibodies (Bethyl) 1:10000 were applied for 1 hour at room temperature in 4% low-fat milk and proteins were visualized using ECL substrate (Thermo Scientific).. Primary antibodies were TSC2 (D93F12) 1:1000, p-Akt S473 (D9E) 1:1000, 4E-BP1 (53H11) 1:1000, p-S6 S240/244 (rabbit polyclonal) 1:3000, S6 (54D2) 1:1000, p-Akt T308 (D25E6) 1:1000, pan-Akt (40D4) 1:1000, β-tubulin (9F3) 1:1000 , p-RB Ser807/811 (D20B12) 1:1000, PDCD4 (D29C6) 1:1000, cleaved caspase 3 Asp175 (5A1E or rabbit polyclonal) 1:500, survivin (71G4B7) 1:1000 and p-S6K1 T389 (108D2) 1:1000, p-TSC2 S939 (rabbit polyclonal) 1:1000 obtained from Cell Signaling Technology.. Bcl-2 (N-19) 1:500, cyclin D1 (M-20) 1:500, p27Kip1 (C-19) 1:1000, cyclin A (C-19) 1:500, CDK4 (C-22) 1:1000, CDK6 (H-96) 1:500, and E2F1 (KH95) 1:500 were fom Santa Cruz. α-tubulin (CP06) 1:3000 was purchased from Calbiochem and GAPDH (rabbit polyclonal) 1:10000 from Trevigen.Bcl-2 (N-19) 1:500, cyclin D1 (M-20) 1:500, p27Kip1 (C-19) 1:1000, cyclin A (C-19) 1:500, CDK4 (C-22) 1:1000, CDK6 (H-96) 1:500, and E2F1 (KH95) 1:500 were fom Santa Cruz. α-tubulin (CP06) 1:3000 was purchased..

    Article Title: TSC1 promotes i NKT cell anergy and inhibits i NKT cell-mediated anti-tumor immunity
    Article Snippet: Fluorescence-conjugated anti-mouse TCRβ (H57-597), NK1.1 (PK136), CD44(IM7), B220(RA3-6B2), Gr-1(RB6-8C5), CD11c(N418), CD11b(M1/70), CD4 (GK1.5), CD8 (53-6.7), IFN-γ (XMG1.2), IL-4 (11B11), PD-1(RMP1-30), and ICOS(D10.G4.1) antibodies were purchased from BioLegend.Fluorescence-conjugated anti-mouse TCRβ (H57-597), NK1.1 (PK136), CD44(IM7), B220(RA3-6B2), Gr-1(RB6-8C5), CD11c(N418), CD11b(M1/70), CD4 (GK1.5), CD8 (53-6.7), IFN-γ (XMG1.2), IL-4 (11B11), PD-1(RMP1-30), and ICOS(D10.G4.1) antibodies were purchased from BioLegend.. Anti–phospho-p44/42 mitogen-activated protein kinase T202/Y204 (197G2), anti–phospho-S6 S240/244 (D68F8), anti–phospho-4E-BP1 T37/46 (236B4), anti–phospho-Akt S473 (193H12), TSC1 (D43E2), and TSC2 (D93F12) were purchased from Cell Signaling Technology, Inc. Anti–β-actin antibody was purchased from Sigma-Aldrich.. Cell surface staining was performed with 2% FBS-PBS.Cell surface staining was performed with 2% FBS-PBS.

    Article Title: TSC1 promotes i NKT cell anergy and inhibits i NKT cell-mediated anti-tumor immunity
    Article Snippet: CD11b(M1/70), CD4 (GK1.5), CD8 (53-6.7), IFN-γ (XMG1.2), IL-4 (11B11), PD-1(RMP1-30), and ICOS(D10.G4.1) antibodies were purchased from BioLegend. .. Anti–phospho-p44/42 mitogen-activated protein kinase T202/Y204 (197G2), anti–phospho-S6 S240/244 (D68F8), anti–phospho-4E-BP1 T37/46 (236B4), anti–phospho-Akt S473 (193H12), TSC1 (D43E2), and TSC2 (D93F12) were purchased from Cell Signaling Technology, Inc. Anti–β-actin antibody was purchased from Sigma-Aldrich. .. Cell surface staining was performed with 2% FBS-PBS.

    Immunofluorescence:

    Article Title: Trans‐gnetin H isolated from the seeds of Paeonia species induces autophagy via inhibiting mTORC1 signalling through AMPK activation
    Article Snippet: The WB protocol was the same as our previous work.The WB protocol was the same as our previous work.. The primary antibodies Actin (20536‐1‐AP), LC3II (18725‐1‐AP), p62 (18420‐1‐AP) were purchased from Proteintech; TFEB (37785 S), pT389‐S6K (9234 S), p‐S6 (4858 S), S6K (9202 S), S6 (2217 S), p‐AMPKα (50081 S), AMPKα (5832 S), Raptor (2280 S) and TSC2 (D93F12) were purchased from Cell Signaling Technology; Flag (db7002) and HA (db2603) were purchased from Hangzhou Bio Technolog.. The secondary antibodies were obtained from Sigma.The secondary antibodies were obtained from Sigma.

    Article Title: A translational program that suppresses metabolism to shield the genome
    Article Snippet: Blots were stripped and reprobed when looking at total versus modified proteins unless otherwise noted.. Antibodies (all 1:1000): GLUT1 (Novus Biologicals, # NB110-39113), PAI-1 (Proteintech, #13801-1-AP), Puromycin (3RH11) (Kerafast, #EQ0001), β-ACTIN (C4) (SantaCruz Biotechnology, #sc-47778), EIF5A (abcam, #ab32443), EIF3D (Proteintech, # 10219-1-AP), EIF3K (Proteintech, # 10640-1-AP), acetyl-EIF5A (Lys47) (Boster Bio, #P01727), CYR61 (Proteintech, #26689-1-AP), SDC4 (R&D Systems, #P31431), SIRT1 (Proteintech, #13161-1-AP), SIRT2 (Proteintech, #19655-1-AP), NFKB2 (Proteintech, #10409-2-AP), 4E-BP (Cell Signaling, #9452 S), Phospho-4E-BP (Ser65) (Cell Signaling, #9451 S), TSC2 (D93F12) (Cell Signaling, #4308S), gamma H2AX (Novus, NB100-384), H2AX (Abcam, Ab11175), CRM-1 (Novus, NB100-79802), XPO-4 (Abcam, ab133237), eIF5A2 (Thermo, #PA5-30770), Phospho-RAPTOR (Ser792) (Cell Signaling, #89146S), RAPTOR (Cell Signaling, #2280), Phospho-Tuberin/TSC2 (Ser1387) (Cell Signaling, #5584S), NDRG1 (Abcam, #ab37897), and Anti-hypusine (mAbHpu24) antibody was kindly provided by Genentech.. The eIF5A1 antibodies and siRNAs used in this study were specific to eIF5A1 and not eIF5A2 as described by the manufacturer and validated in Supplementary Fig. .

    Article Title: Identification of mechanically regulated phosphorylation sites on tuberin (TSC2) that control mechanistic target of rapamycin (mTOR) signaling
    Article Snippet: Images were quantified using ImageJ software (U.S. National Institutes of Health, Bethesda, MD; http://rsb.info.nih.gov/nih-image/).Images were quantified using ImageJ software (U.S. National Institutes of Health, Bethesda, MD; http://rsb.info.nih.gov/nih-image/).. Antibodies - P-p70(389) (1A5) (#9206, used to probe Myc-tagged P-p70), P-p70(389) (#9234, used to probe endogenous P-p70), total p70 (49D7) (#2708), Anti-P-Akt substrate (RxRxxS*/T*) (23C8D2) for detection of PTSC2 (#10001), P-JNK(Thr183/Tyr185) (98F2) (#4671), total JNK (#9252), total mTOR (#2972), total Rictor (D16H9) (#9476), P-PKB (Ser473) (#4060), P-PKB (Thr308) (#9275S), total PKB (#9272), P-S6 (Ser240/4) (#5364), P-S6 (Ser235/6) (#2211), total S6 (#2217S), total TSC1 (D43E2) (#6935), total TSC2 (D93F12) (#4308) were purchased from Cell Signaling Technology (Danvers, MA, USA).. Anti-puromycin (12D10) (#MABE343) was purchased from EMD Millipore (Billerica, MA, USA).Anti-puromycin (12D10) (#MABE343) was purchased from EMD Millipore (Billerica, MA, USA).

    Article Title: Chronic signaling via the metabolic checkpoint kinase mTORC1 induces macrophage granuloma formation and marks sarcoidosis progression
    Article Snippet: HRP-conjugated secondary antibodies (Bethyl) 1:10000 were applied for 1 hour at room temperature in 4% low-fat milk and proteins were visualized using ECL substrate (Thermo Scientific).HRP-conjugated secondary antibodies (Bethyl) 1:10000 were applied for 1 hour at room temperature in 4% low-fat milk and proteins were visualized using ECL substrate (Thermo Scientific).. Primary antibodies were TSC2 (D93F12) 1:1000, p-Akt S473 (D9E) 1:1000, 4E-BP1 (53H11) 1:1000, p-S6 S240/244 (rabbit polyclonal) 1:3000, S6 (54D2) 1:1000, p-Akt T308 (D25E6) 1:1000, pan-Akt (40D4) 1:1000, β-tubulin (9F3) 1:1000 , p-RB Ser807/811 (D20B12) 1:1000, PDCD4 (D29C6) 1:1000, cleaved caspase 3 Asp175 (5A1E or rabbit polyclonal) 1:500, survivin (71G4B7) 1:1000 and p-S6K1 T389 (108D2) 1:1000, p-TSC2 S939 (rabbit polyclonal) 1:1000 obtained from Cell Signaling Technology.. Bcl-2 (N-19) 1:500, cyclin D1 (M-20) 1:500, p27Kip1 (C-19) 1:1000, cyclin A (C-19) 1:500, CDK4 (C-22) 1:1000, CDK6 (H-96) 1:500, and E2F1 (KH95) 1:500 were fom Santa Cruz. α-tubulin (CP06) 1:3000 was purchased from Calbiochem and GAPDH (rabbit polyclonal) 1:10000 from Trevigen.Bcl-2 (N-19) 1:500, cyclin D1 (M-20) 1:500, p27Kip1 (C-19) 1:1000, cyclin A (C-19) 1:500, CDK4 (C-22) 1:1000, CDK6 (H-96) 1:500, and E2F1 (KH95) 1:500 were fom Santa Cruz. α-tubulin (CP06) 1:3000 was purchased..

    Article Title: TSC1 promotes i NKT cell anergy and inhibits i NKT cell-mediated anti-tumor immunity
    Article Snippet: Fluorescence-conjugated anti-mouse TCRβ (H57-597), NK1.1 (PK136), CD44(IM7), B220(RA3-6B2), Gr-1(RB6-8C5), CD11c(N418), CD11b(M1/70), CD4 (GK1.5), CD8 (53-6.7), IFN-γ (XMG1.2), IL-4 (11B11), PD-1(RMP1-30), and ICOS(D10.G4.1) antibodies were purchased from BioLegend.Fluorescence-conjugated anti-mouse TCRβ (H57-597), NK1.1 (PK136), CD44(IM7), B220(RA3-6B2), Gr-1(RB6-8C5), CD11c(N418), CD11b(M1/70), CD4 (GK1.5), CD8 (53-6.7), IFN-γ (XMG1.2), IL-4 (11B11), PD-1(RMP1-30), and ICOS(D10.G4.1) antibodies were purchased from BioLegend.. Anti–phospho-p44/42 mitogen-activated protein kinase T202/Y204 (197G2), anti–phospho-S6 S240/244 (D68F8), anti–phospho-4E-BP1 T37/46 (236B4), anti–phospho-Akt S473 (193H12), TSC1 (D43E2), and TSC2 (D93F12) were purchased from Cell Signaling Technology, Inc. Anti–β-actin antibody was purchased from Sigma-Aldrich.. Cell surface staining was performed with 2% FBS-PBS.Cell surface staining was performed with 2% FBS-PBS.

    Article Title: TSC1 promotes i NKT cell anergy and inhibits i NKT cell-mediated anti-tumor immunity
    Article Snippet: CD11b(M1/70), CD4 (GK1.5), CD8 (53-6.7), IFN-γ (XMG1.2), IL-4 (11B11), PD-1(RMP1-30), and ICOS(D10.G4.1) antibodies were purchased from BioLegend. .. Anti–phospho-p44/42 mitogen-activated protein kinase T202/Y204 (197G2), anti–phospho-S6 S240/244 (D68F8), anti–phospho-4E-BP1 T37/46 (236B4), anti–phospho-Akt S473 (193H12), TSC1 (D43E2), and TSC2 (D93F12) were purchased from Cell Signaling Technology, Inc. Anti–β-actin antibody was purchased from Sigma-Aldrich. .. Cell surface staining was performed with 2% FBS-PBS.

    Phospho-proteomics:

    Article Title: Trans‐gnetin H isolated from the seeds of Paeonia species induces autophagy via inhibiting mTORC1 signalling through AMPK activation
    Article Snippet: The WB protocol was the same as our previous work.The WB protocol was the same as our previous work.. The primary antibodies Actin (20536‐1‐AP), LC3II (18725‐1‐AP), p62 (18420‐1‐AP) were purchased from Proteintech; TFEB (37785 S), pT389‐S6K (9234 S), p‐S6 (4858 S), S6K (9202 S), S6 (2217 S), p‐AMPKα (50081 S), AMPKα (5832 S), Raptor (2280 S) and TSC2 (D93F12) were purchased from Cell Signaling Technology; Flag (db7002) and HA (db2603) were purchased from Hangzhou Bio Technolog.. The secondary antibodies were obtained from Sigma.The secondary antibodies were obtained from Sigma.

    Article Title: A translational program that suppresses metabolism to shield the genome
    Article Snippet: Blots were stripped and reprobed when looking at total versus modified proteins unless otherwise noted.. Antibodies (all 1:1000): GLUT1 (Novus Biologicals, # NB110-39113), PAI-1 (Proteintech, #13801-1-AP), Puromycin (3RH11) (Kerafast, #EQ0001), β-ACTIN (C4) (SantaCruz Biotechnology, #sc-47778), EIF5A (abcam, #ab32443), EIF3D (Proteintech, # 10219-1-AP), EIF3K (Proteintech, # 10640-1-AP), acetyl-EIF5A (Lys47) (Boster Bio, #P01727), CYR61 (Proteintech, #26689-1-AP), SDC4 (R&D Systems, #P31431), SIRT1 (Proteintech, #13161-1-AP), SIRT2 (Proteintech, #19655-1-AP), NFKB2 (Proteintech, #10409-2-AP), 4E-BP (Cell Signaling, #9452 S), Phospho-4E-BP (Ser65) (Cell Signaling, #9451 S), TSC2 (D93F12) (Cell Signaling, #4308S), gamma H2AX (Novus, NB100-384), H2AX (Abcam, Ab11175), CRM-1 (Novus, NB100-79802), XPO-4 (Abcam, ab133237), eIF5A2 (Thermo, #PA5-30770), Phospho-RAPTOR (Ser792) (Cell Signaling, #89146S), RAPTOR (Cell Signaling, #2280), Phospho-Tuberin/TSC2 (Ser1387) (Cell Signaling, #5584S), NDRG1 (Abcam, #ab37897), and Anti-hypusine (mAbHpu24) antibody was kindly provided by Genentech.. The eIF5A1 antibodies and siRNAs used in this study were specific to eIF5A1 and not eIF5A2 as described by the manufacturer and validated in Supplementary Fig. .

    Article Title: Identification of mechanically regulated phosphorylation sites on tuberin (TSC2) that control mechanistic target of rapamycin (mTOR) signaling
    Article Snippet: Images were quantified using ImageJ software (U.S. National Institutes of Health, Bethesda, MD; http://rsb.info.nih.gov/nih-image/).Images were quantified using ImageJ software (U.S. National Institutes of Health, Bethesda, MD; http://rsb.info.nih.gov/nih-image/).. Antibodies - P-p70(389) (1A5) (#9206, used to probe Myc-tagged P-p70), P-p70(389) (#9234, used to probe endogenous P-p70), total p70 (49D7) (#2708), Anti-P-Akt substrate (RxRxxS*/T*) (23C8D2) for detection of PTSC2 (#10001), P-JNK(Thr183/Tyr185) (98F2) (#4671), total JNK (#9252), total mTOR (#2972), total Rictor (D16H9) (#9476), P-PKB (Ser473) (#4060), P-PKB (Thr308) (#9275S), total PKB (#9272), P-S6 (Ser240/4) (#5364), P-S6 (Ser235/6) (#2211), total S6 (#2217S), total TSC1 (D43E2) (#6935), total TSC2 (D93F12) (#4308) were purchased from Cell Signaling Technology (Danvers, MA, USA).. Anti-puromycin (12D10) (#MABE343) was purchased from EMD Millipore (Billerica, MA, USA).Anti-puromycin (12D10) (#MABE343) was purchased from EMD Millipore (Billerica, MA, USA).

    Article Title: Chronic signaling via the metabolic checkpoint kinase mTORC1 induces macrophage granuloma formation and marks sarcoidosis progression
    Article Snippet: HRP-conjugated secondary antibodies (Bethyl) 1:10000 were applied for 1 hour at room temperature in 4% low-fat milk and proteins were visualized using ECL substrate (Thermo Scientific).HRP-conjugated secondary antibodies (Bethyl) 1:10000 were applied for 1 hour at room temperature in 4% low-fat milk and proteins were visualized using ECL substrate (Thermo Scientific).. Primary antibodies were TSC2 (D93F12) 1:1000, p-Akt S473 (D9E) 1:1000, 4E-BP1 (53H11) 1:1000, p-S6 S240/244 (rabbit polyclonal) 1:3000, S6 (54D2) 1:1000, p-Akt T308 (D25E6) 1:1000, pan-Akt (40D4) 1:1000, β-tubulin (9F3) 1:1000 , p-RB Ser807/811 (D20B12) 1:1000, PDCD4 (D29C6) 1:1000, cleaved caspase 3 Asp175 (5A1E or rabbit polyclonal) 1:500, survivin (71G4B7) 1:1000 and p-S6K1 T389 (108D2) 1:1000, p-TSC2 S939 (rabbit polyclonal) 1:1000 obtained from Cell Signaling Technology.. Bcl-2 (N-19) 1:500, cyclin D1 (M-20) 1:500, p27Kip1 (C-19) 1:1000, cyclin A (C-19) 1:500, CDK4 (C-22) 1:1000, CDK6 (H-96) 1:500, and E2F1 (KH95) 1:500 were fom Santa Cruz. α-tubulin (CP06) 1:3000 was purchased from Calbiochem and GAPDH (rabbit polyclonal) 1:10000 from Trevigen.Bcl-2 (N-19) 1:500, cyclin D1 (M-20) 1:500, p27Kip1 (C-19) 1:1000, cyclin A (C-19) 1:500, CDK4 (C-22) 1:1000, CDK6 (H-96) 1:500, and E2F1 (KH95) 1:500 were fom Santa Cruz. α-tubulin (CP06) 1:3000 was purchased..

    Article Title: TSC1 promotes i NKT cell anergy and inhibits i NKT cell-mediated anti-tumor immunity
    Article Snippet: Fluorescence-conjugated anti-mouse TCRβ (H57-597), NK1.1 (PK136), CD44(IM7), B220(RA3-6B2), Gr-1(RB6-8C5), CD11c(N418), CD11b(M1/70), CD4 (GK1.5), CD8 (53-6.7), IFN-γ (XMG1.2), IL-4 (11B11), PD-1(RMP1-30), and ICOS(D10.G4.1) antibodies were purchased from BioLegend.Fluorescence-conjugated anti-mouse TCRβ (H57-597), NK1.1 (PK136), CD44(IM7), B220(RA3-6B2), Gr-1(RB6-8C5), CD11c(N418), CD11b(M1/70), CD4 (GK1.5), CD8 (53-6.7), IFN-γ (XMG1.2), IL-4 (11B11), PD-1(RMP1-30), and ICOS(D10.G4.1) antibodies were purchased from BioLegend.. Anti–phospho-p44/42 mitogen-activated protein kinase T202/Y204 (197G2), anti–phospho-S6 S240/244 (D68F8), anti–phospho-4E-BP1 T37/46 (236B4), anti–phospho-Akt S473 (193H12), TSC1 (D43E2), and TSC2 (D93F12) were purchased from Cell Signaling Technology, Inc. Anti–β-actin antibody was purchased from Sigma-Aldrich.. Cell surface staining was performed with 2% FBS-PBS.Cell surface staining was performed with 2% FBS-PBS.

    Article Title: TSC1 promotes i NKT cell anergy and inhibits i NKT cell-mediated anti-tumor immunity
    Article Snippet: CD11b(M1/70), CD4 (GK1.5), CD8 (53-6.7), IFN-γ (XMG1.2), IL-4 (11B11), PD-1(RMP1-30), and ICOS(D10.G4.1) antibodies were purchased from BioLegend. .. Anti–phospho-p44/42 mitogen-activated protein kinase T202/Y204 (197G2), anti–phospho-S6 S240/244 (D68F8), anti–phospho-4E-BP1 T37/46 (236B4), anti–phospho-Akt S473 (193H12), TSC1 (D43E2), and TSC2 (D93F12) were purchased from Cell Signaling Technology, Inc. Anti–β-actin antibody was purchased from Sigma-Aldrich. .. Cell surface staining was performed with 2% FBS-PBS.

    Knockdown:

    Article Title: Trans‐gnetin H isolated from the seeds of Paeonia species induces autophagy via inhibiting mTORC1 signalling through AMPK activation
    Article Snippet: The WB protocol was the same as our previous work.The WB protocol was the same as our previous work.. The primary antibodies Actin (20536‐1‐AP), LC3II (18725‐1‐AP), p62 (18420‐1‐AP) were purchased from Proteintech; TFEB (37785 S), pT389‐S6K (9234 S), p‐S6 (4858 S), S6K (9202 S), S6 (2217 S), p‐AMPKα (50081 S), AMPKα (5832 S), Raptor (2280 S) and TSC2 (D93F12) were purchased from Cell Signaling Technology; Flag (db7002) and HA (db2603) were purchased from Hangzhou Bio Technolog.. The secondary antibodies were obtained from Sigma.The secondary antibodies were obtained from Sigma.

    Article Title: A translational program that suppresses metabolism to shield the genome
    Article Snippet: Blots were stripped and reprobed when looking at total versus modified proteins unless otherwise noted.. Antibodies (all 1:1000): GLUT1 (Novus Biologicals, # NB110-39113), PAI-1 (Proteintech, #13801-1-AP), Puromycin (3RH11) (Kerafast, #EQ0001), β-ACTIN (C4) (SantaCruz Biotechnology, #sc-47778), EIF5A (abcam, #ab32443), EIF3D (Proteintech, # 10219-1-AP), EIF3K (Proteintech, # 10640-1-AP), acetyl-EIF5A (Lys47) (Boster Bio, #P01727), CYR61 (Proteintech, #26689-1-AP), SDC4 (R&D Systems, #P31431), SIRT1 (Proteintech, #13161-1-AP), SIRT2 (Proteintech, #19655-1-AP), NFKB2 (Proteintech, #10409-2-AP), 4E-BP (Cell Signaling, #9452 S), Phospho-4E-BP (Ser65) (Cell Signaling, #9451 S), TSC2 (D93F12) (Cell Signaling, #4308S), gamma H2AX (Novus, NB100-384), H2AX (Abcam, Ab11175), CRM-1 (Novus, NB100-79802), XPO-4 (Abcam, ab133237), eIF5A2 (Thermo, #PA5-30770), Phospho-RAPTOR (Ser792) (Cell Signaling, #89146S), RAPTOR (Cell Signaling, #2280), Phospho-Tuberin/TSC2 (Ser1387) (Cell Signaling, #5584S), NDRG1 (Abcam, #ab37897), and Anti-hypusine (mAbHpu24) antibody was kindly provided by Genentech.. The eIF5A1 antibodies and siRNAs used in this study were specific to eIF5A1 and not eIF5A2 as described by the manufacturer and validated in Supplementary Fig. .

    Article Title: Identification of mechanically regulated phosphorylation sites on tuberin (TSC2) that control mechanistic target of rapamycin (mTOR) signaling
    Article Snippet: Images were quantified using ImageJ software (U.S. National Institutes of Health, Bethesda, MD; http://rsb.info.nih.gov/nih-image/).Images were quantified using ImageJ software (U.S. National Institutes of Health, Bethesda, MD; http://rsb.info.nih.gov/nih-image/).. Antibodies - P-p70(389) (1A5) (#9206, used to probe Myc-tagged P-p70), P-p70(389) (#9234, used to probe endogenous P-p70), total p70 (49D7) (#2708), Anti-P-Akt substrate (RxRxxS*/T*) (23C8D2) for detection of PTSC2 (#10001), P-JNK(Thr183/Tyr185) (98F2) (#4671), total JNK (#9252), total mTOR (#2972), total Rictor (D16H9) (#9476), P-PKB (Ser473) (#4060), P-PKB (Thr308) (#9275S), total PKB (#9272), P-S6 (Ser240/4) (#5364), P-S6 (Ser235/6) (#2211), total S6 (#2217S), total TSC1 (D43E2) (#6935), total TSC2 (D93F12) (#4308) were purchased from Cell Signaling Technology (Danvers, MA, USA).. Anti-puromycin (12D10) (#MABE343) was purchased from EMD Millipore (Billerica, MA, USA).Anti-puromycin (12D10) (#MABE343) was purchased from EMD Millipore (Billerica, MA, USA).

    Article Title: Chronic signaling via the metabolic checkpoint kinase mTORC1 induces macrophage granuloma formation and marks sarcoidosis progression
    Article Snippet: HRP-conjugated secondary antibodies (Bethyl) 1:10000 were applied for 1 hour at room temperature in 4% low-fat milk and proteins were visualized using ECL substrate (Thermo Scientific).HRP-conjugated secondary antibodies (Bethyl) 1:10000 were applied for 1 hour at room temperature in 4% low-fat milk and proteins were visualized using ECL substrate (Thermo Scientific).. Primary antibodies were TSC2 (D93F12) 1:1000, p-Akt S473 (D9E) 1:1000, 4E-BP1 (53H11) 1:1000, p-S6 S240/244 (rabbit polyclonal) 1:3000, S6 (54D2) 1:1000, p-Akt T308 (D25E6) 1:1000, pan-Akt (40D4) 1:1000, β-tubulin (9F3) 1:1000 , p-RB Ser807/811 (D20B12) 1:1000, PDCD4 (D29C6) 1:1000, cleaved caspase 3 Asp175 (5A1E or rabbit polyclonal) 1:500, survivin (71G4B7) 1:1000 and p-S6K1 T389 (108D2) 1:1000, p-TSC2 S939 (rabbit polyclonal) 1:1000 obtained from Cell Signaling Technology.. Bcl-2 (N-19) 1:500, cyclin D1 (M-20) 1:500, p27Kip1 (C-19) 1:1000, cyclin A (C-19) 1:500, CDK4 (C-22) 1:1000, CDK6 (H-96) 1:500, and E2F1 (KH95) 1:500 were fom Santa Cruz. α-tubulin (CP06) 1:3000 was purchased from Calbiochem and GAPDH (rabbit polyclonal) 1:10000 from Trevigen.Bcl-2 (N-19) 1:500, cyclin D1 (M-20) 1:500, p27Kip1 (C-19) 1:1000, cyclin A (C-19) 1:500, CDK4 (C-22) 1:1000, CDK6 (H-96) 1:500, and E2F1 (KH95) 1:500 were fom Santa Cruz. α-tubulin (CP06) 1:3000 was purchased..

    Article Title: TSC1 promotes i NKT cell anergy and inhibits i NKT cell-mediated anti-tumor immunity
    Article Snippet: Fluorescence-conjugated anti-mouse TCRβ (H57-597), NK1.1 (PK136), CD44(IM7), B220(RA3-6B2), Gr-1(RB6-8C5), CD11c(N418), CD11b(M1/70), CD4 (GK1.5), CD8 (53-6.7), IFN-γ (XMG1.2), IL-4 (11B11), PD-1(RMP1-30), and ICOS(D10.G4.1) antibodies were purchased from BioLegend.Fluorescence-conjugated anti-mouse TCRβ (H57-597), NK1.1 (PK136), CD44(IM7), B220(RA3-6B2), Gr-1(RB6-8C5), CD11c(N418), CD11b(M1/70), CD4 (GK1.5), CD8 (53-6.7), IFN-γ (XMG1.2), IL-4 (11B11), PD-1(RMP1-30), and ICOS(D10.G4.1) antibodies were purchased from BioLegend.. Anti–phospho-p44/42 mitogen-activated protein kinase T202/Y204 (197G2), anti–phospho-S6 S240/244 (D68F8), anti–phospho-4E-BP1 T37/46 (236B4), anti–phospho-Akt S473 (193H12), TSC1 (D43E2), and TSC2 (D93F12) were purchased from Cell Signaling Technology, Inc. Anti–β-actin antibody was purchased from Sigma-Aldrich.. Cell surface staining was performed with 2% FBS-PBS.Cell surface staining was performed with 2% FBS-PBS.

    Article Title: TSC1 promotes i NKT cell anergy and inhibits i NKT cell-mediated anti-tumor immunity
    Article Snippet: CD11b(M1/70), CD4 (GK1.5), CD8 (53-6.7), IFN-γ (XMG1.2), IL-4 (11B11), PD-1(RMP1-30), and ICOS(D10.G4.1) antibodies were purchased from BioLegend. .. Anti–phospho-p44/42 mitogen-activated protein kinase T202/Y204 (197G2), anti–phospho-S6 S240/244 (D68F8), anti–phospho-4E-BP1 T37/46 (236B4), anti–phospho-Akt S473 (193H12), TSC1 (D43E2), and TSC2 (D93F12) were purchased from Cell Signaling Technology, Inc. Anti–β-actin antibody was purchased from Sigma-Aldrich. .. Cell surface staining was performed with 2% FBS-PBS.

    Co-Immunoprecipitation Assay:

    Article Title: Trans‐gnetin H isolated from the seeds of Paeonia species induces autophagy via inhibiting mTORC1 signalling through AMPK activation
    Article Snippet: The WB protocol was the same as our previous work.The WB protocol was the same as our previous work.. The primary antibodies Actin (20536‐1‐AP), LC3II (18725‐1‐AP), p62 (18420‐1‐AP) were purchased from Proteintech; TFEB (37785 S), pT389‐S6K (9234 S), p‐S6 (4858 S), S6K (9202 S), S6 (2217 S), p‐AMPKα (50081 S), AMPKα (5832 S), Raptor (2280 S) and TSC2 (D93F12) were purchased from Cell Signaling Technology; Flag (db7002) and HA (db2603) were purchased from Hangzhou Bio Technolog.. The secondary antibodies were obtained from Sigma.The secondary antibodies were obtained from Sigma.

    Article Title: A translational program that suppresses metabolism to shield the genome
    Article Snippet: Blots were stripped and reprobed when looking at total versus modified proteins unless otherwise noted.. Antibodies (all 1:1000): GLUT1 (Novus Biologicals, # NB110-39113), PAI-1 (Proteintech, #13801-1-AP), Puromycin (3RH11) (Kerafast, #EQ0001), β-ACTIN (C4) (SantaCruz Biotechnology, #sc-47778), EIF5A (abcam, #ab32443), EIF3D (Proteintech, # 10219-1-AP), EIF3K (Proteintech, # 10640-1-AP), acetyl-EIF5A (Lys47) (Boster Bio, #P01727), CYR61 (Proteintech, #26689-1-AP), SDC4 (R&D Systems, #P31431), SIRT1 (Proteintech, #13161-1-AP), SIRT2 (Proteintech, #19655-1-AP), NFKB2 (Proteintech, #10409-2-AP), 4E-BP (Cell Signaling, #9452 S), Phospho-4E-BP (Ser65) (Cell Signaling, #9451 S), TSC2 (D93F12) (Cell Signaling, #4308S), gamma H2AX (Novus, NB100-384), H2AX (Abcam, Ab11175), CRM-1 (Novus, NB100-79802), XPO-4 (Abcam, ab133237), eIF5A2 (Thermo, #PA5-30770), Phospho-RAPTOR (Ser792) (Cell Signaling, #89146S), RAPTOR (Cell Signaling, #2280), Phospho-Tuberin/TSC2 (Ser1387) (Cell Signaling, #5584S), NDRG1 (Abcam, #ab37897), and Anti-hypusine (mAbHpu24) antibody was kindly provided by Genentech.. The eIF5A1 antibodies and siRNAs used in this study were specific to eIF5A1 and not eIF5A2 as described by the manufacturer and validated in Supplementary Fig. .

    Article Title: Identification of mechanically regulated phosphorylation sites on tuberin (TSC2) that control mechanistic target of rapamycin (mTOR) signaling
    Article Snippet: Images were quantified using ImageJ software (U.S. National Institutes of Health, Bethesda, MD; http://rsb.info.nih.gov/nih-image/).Images were quantified using ImageJ software (U.S. National Institutes of Health, Bethesda, MD; http://rsb.info.nih.gov/nih-image/).. Antibodies - P-p70(389) (1A5) (#9206, used to probe Myc-tagged P-p70), P-p70(389) (#9234, used to probe endogenous P-p70), total p70 (49D7) (#2708), Anti-P-Akt substrate (RxRxxS*/T*) (23C8D2) for detection of PTSC2 (#10001), P-JNK(Thr183/Tyr185) (98F2) (#4671), total JNK (#9252), total mTOR (#2972), total Rictor (D16H9) (#9476), P-PKB (Ser473) (#4060), P-PKB (Thr308) (#9275S), total PKB (#9272), P-S6 (Ser240/4) (#5364), P-S6 (Ser235/6) (#2211), total S6 (#2217S), total TSC1 (D43E2) (#6935), total TSC2 (D93F12) (#4308) were purchased from Cell Signaling Technology (Danvers, MA, USA).. Anti-puromycin (12D10) (#MABE343) was purchased from EMD Millipore (Billerica, MA, USA).Anti-puromycin (12D10) (#MABE343) was purchased from EMD Millipore (Billerica, MA, USA).

    Article Title: Chronic signaling via the metabolic checkpoint kinase mTORC1 induces macrophage granuloma formation and marks sarcoidosis progression
    Article Snippet: HRP-conjugated secondary antibodies (Bethyl) 1:10000 were applied for 1 hour at room temperature in 4% low-fat milk and proteins were visualized using ECL substrate (Thermo Scientific).HRP-conjugated secondary antibodies (Bethyl) 1:10000 were applied for 1 hour at room temperature in 4% low-fat milk and proteins were visualized using ECL substrate (Thermo Scientific).. Primary antibodies were TSC2 (D93F12) 1:1000, p-Akt S473 (D9E) 1:1000, 4E-BP1 (53H11) 1:1000, p-S6 S240/244 (rabbit polyclonal) 1:3000, S6 (54D2) 1:1000, p-Akt T308 (D25E6) 1:1000, pan-Akt (40D4) 1:1000, β-tubulin (9F3) 1:1000 , p-RB Ser807/811 (D20B12) 1:1000, PDCD4 (D29C6) 1:1000, cleaved caspase 3 Asp175 (5A1E or rabbit polyclonal) 1:500, survivin (71G4B7) 1:1000 and p-S6K1 T389 (108D2) 1:1000, p-TSC2 S939 (rabbit polyclonal) 1:1000 obtained from Cell Signaling Technology.. Bcl-2 (N-19) 1:500, cyclin D1 (M-20) 1:500, p27Kip1 (C-19) 1:1000, cyclin A (C-19) 1:500, CDK4 (C-22) 1:1000, CDK6 (H-96) 1:500, and E2F1 (KH95) 1:500 were fom Santa Cruz. α-tubulin (CP06) 1:3000 was purchased from Calbiochem and GAPDH (rabbit polyclonal) 1:10000 from Trevigen.Bcl-2 (N-19) 1:500, cyclin D1 (M-20) 1:500, p27Kip1 (C-19) 1:1000, cyclin A (C-19) 1:500, CDK4 (C-22) 1:1000, CDK6 (H-96) 1:500, and E2F1 (KH95) 1:500 were fom Santa Cruz. α-tubulin (CP06) 1:3000 was purchased..

    Article Title: TSC1 promotes i NKT cell anergy and inhibits i NKT cell-mediated anti-tumor immunity
    Article Snippet: Fluorescence-conjugated anti-mouse TCRβ (H57-597), NK1.1 (PK136), CD44(IM7), B220(RA3-6B2), Gr-1(RB6-8C5), CD11c(N418), CD11b(M1/70), CD4 (GK1.5), CD8 (53-6.7), IFN-γ (XMG1.2), IL-4 (11B11), PD-1(RMP1-30), and ICOS(D10.G4.1) antibodies were purchased from BioLegend.Fluorescence-conjugated anti-mouse TCRβ (H57-597), NK1.1 (PK136), CD44(IM7), B220(RA3-6B2), Gr-1(RB6-8C5), CD11c(N418), CD11b(M1/70), CD4 (GK1.5), CD8 (53-6.7), IFN-γ (XMG1.2), IL-4 (11B11), PD-1(RMP1-30), and ICOS(D10.G4.1) antibodies were purchased from BioLegend.. Anti–phospho-p44/42 mitogen-activated protein kinase T202/Y204 (197G2), anti–phospho-S6 S240/244 (D68F8), anti–phospho-4E-BP1 T37/46 (236B4), anti–phospho-Akt S473 (193H12), TSC1 (D43E2), and TSC2 (D93F12) were purchased from Cell Signaling Technology, Inc. Anti–β-actin antibody was purchased from Sigma-Aldrich.. Cell surface staining was performed with 2% FBS-PBS.Cell surface staining was performed with 2% FBS-PBS.

    Article Title: TSC1 promotes i NKT cell anergy and inhibits i NKT cell-mediated anti-tumor immunity
    Article Snippet: CD11b(M1/70), CD4 (GK1.5), CD8 (53-6.7), IFN-γ (XMG1.2), IL-4 (11B11), PD-1(RMP1-30), and ICOS(D10.G4.1) antibodies were purchased from BioLegend. .. Anti–phospho-p44/42 mitogen-activated protein kinase T202/Y204 (197G2), anti–phospho-S6 S240/244 (D68F8), anti–phospho-4E-BP1 T37/46 (236B4), anti–phospho-Akt S473 (193H12), TSC1 (D43E2), and TSC2 (D93F12) were purchased from Cell Signaling Technology, Inc. Anti–β-actin antibody was purchased from Sigma-Aldrich. .. Cell surface staining was performed with 2% FBS-PBS.

    Expressing:

    Article Title: Trans‐gnetin H isolated from the seeds of Paeonia species induces autophagy via inhibiting mTORC1 signalling through AMPK activation
    Article Snippet: The WB protocol was the same as our previous work.The WB protocol was the same as our previous work.. The primary antibodies Actin (20536‐1‐AP), LC3II (18725‐1‐AP), p62 (18420‐1‐AP) were purchased from Proteintech; TFEB (37785 S), pT389‐S6K (9234 S), p‐S6 (4858 S), S6K (9202 S), S6 (2217 S), p‐AMPKα (50081 S), AMPKα (5832 S), Raptor (2280 S) and TSC2 (D93F12) were purchased from Cell Signaling Technology; Flag (db7002) and HA (db2603) were purchased from Hangzhou Bio Technolog.. The secondary antibodies were obtained from Sigma.The secondary antibodies were obtained from Sigma.

    Article Title: A translational program that suppresses metabolism to shield the genome
    Article Snippet: Blots were stripped and reprobed when looking at total versus modified proteins unless otherwise noted.. Antibodies (all 1:1000): GLUT1 (Novus Biologicals, # NB110-39113), PAI-1 (Proteintech, #13801-1-AP), Puromycin (3RH11) (Kerafast, #EQ0001), β-ACTIN (C4) (SantaCruz Biotechnology, #sc-47778), EIF5A (abcam, #ab32443), EIF3D (Proteintech, # 10219-1-AP), EIF3K (Proteintech, # 10640-1-AP), acetyl-EIF5A (Lys47) (Boster Bio, #P01727), CYR61 (Proteintech, #26689-1-AP), SDC4 (R&D Systems, #P31431), SIRT1 (Proteintech, #13161-1-AP), SIRT2 (Proteintech, #19655-1-AP), NFKB2 (Proteintech, #10409-2-AP), 4E-BP (Cell Signaling, #9452 S), Phospho-4E-BP (Ser65) (Cell Signaling, #9451 S), TSC2 (D93F12) (Cell Signaling, #4308S), gamma H2AX (Novus, NB100-384), H2AX (Abcam, Ab11175), CRM-1 (Novus, NB100-79802), XPO-4 (Abcam, ab133237), eIF5A2 (Thermo, #PA5-30770), Phospho-RAPTOR (Ser792) (Cell Signaling, #89146S), RAPTOR (Cell Signaling, #2280), Phospho-Tuberin/TSC2 (Ser1387) (Cell Signaling, #5584S), NDRG1 (Abcam, #ab37897), and Anti-hypusine (mAbHpu24) antibody was kindly provided by Genentech.. The eIF5A1 antibodies and siRNAs used in this study were specific to eIF5A1 and not eIF5A2 as described by the manufacturer and validated in Supplementary Fig. .

    Article Title: Identification of mechanically regulated phosphorylation sites on tuberin (TSC2) that control mechanistic target of rapamycin (mTOR) signaling
    Article Snippet: Images were quantified using ImageJ software (U.S. National Institutes of Health, Bethesda, MD; http://rsb.info.nih.gov/nih-image/).Images were quantified using ImageJ software (U.S. National Institutes of Health, Bethesda, MD; http://rsb.info.nih.gov/nih-image/).. Antibodies - P-p70(389) (1A5) (#9206, used to probe Myc-tagged P-p70), P-p70(389) (#9234, used to probe endogenous P-p70), total p70 (49D7) (#2708), Anti-P-Akt substrate (RxRxxS*/T*) (23C8D2) for detection of PTSC2 (#10001), P-JNK(Thr183/Tyr185) (98F2) (#4671), total JNK (#9252), total mTOR (#2972), total Rictor (D16H9) (#9476), P-PKB (Ser473) (#4060), P-PKB (Thr308) (#9275S), total PKB (#9272), P-S6 (Ser240/4) (#5364), P-S6 (Ser235/6) (#2211), total S6 (#2217S), total TSC1 (D43E2) (#6935), total TSC2 (D93F12) (#4308) were purchased from Cell Signaling Technology (Danvers, MA, USA).. Anti-puromycin (12D10) (#MABE343) was purchased from EMD Millipore (Billerica, MA, USA).Anti-puromycin (12D10) (#MABE343) was purchased from EMD Millipore (Billerica, MA, USA).

    Article Title: Chronic signaling via the metabolic checkpoint kinase mTORC1 induces macrophage granuloma formation and marks sarcoidosis progression
    Article Snippet: HRP-conjugated secondary antibodies (Bethyl) 1:10000 were applied for 1 hour at room temperature in 4% low-fat milk and proteins were visualized using ECL substrate (Thermo Scientific).HRP-conjugated secondary antibodies (Bethyl) 1:10000 were applied for 1 hour at room temperature in 4% low-fat milk and proteins were visualized using ECL substrate (Thermo Scientific).. Primary antibodies were TSC2 (D93F12) 1:1000, p-Akt S473 (D9E) 1:1000, 4E-BP1 (53H11) 1:1000, p-S6 S240/244 (rabbit polyclonal) 1:3000, S6 (54D2) 1:1000, p-Akt T308 (D25E6) 1:1000, pan-Akt (40D4) 1:1000, β-tubulin (9F3) 1:1000 , p-RB Ser807/811 (D20B12) 1:1000, PDCD4 (D29C6) 1:1000, cleaved caspase 3 Asp175 (5A1E or rabbit polyclonal) 1:500, survivin (71G4B7) 1:1000 and p-S6K1 T389 (108D2) 1:1000, p-TSC2 S939 (rabbit polyclonal) 1:1000 obtained from Cell Signaling Technology.. Bcl-2 (N-19) 1:500, cyclin D1 (M-20) 1:500, p27Kip1 (C-19) 1:1000, cyclin A (C-19) 1:500, CDK4 (C-22) 1:1000, CDK6 (H-96) 1:500, and E2F1 (KH95) 1:500 were fom Santa Cruz. α-tubulin (CP06) 1:3000 was purchased from Calbiochem and GAPDH (rabbit polyclonal) 1:10000 from Trevigen.Bcl-2 (N-19) 1:500, cyclin D1 (M-20) 1:500, p27Kip1 (C-19) 1:1000, cyclin A (C-19) 1:500, CDK4 (C-22) 1:1000, CDK6 (H-96) 1:500, and E2F1 (KH95) 1:500 were fom Santa Cruz. α-tubulin (CP06) 1:3000 was purchased..

    Article Title: TSC1 promotes i NKT cell anergy and inhibits i NKT cell-mediated anti-tumor immunity
    Article Snippet: Fluorescence-conjugated anti-mouse TCRβ (H57-597), NK1.1 (PK136), CD44(IM7), B220(RA3-6B2), Gr-1(RB6-8C5), CD11c(N418), CD11b(M1/70), CD4 (GK1.5), CD8 (53-6.7), IFN-γ (XMG1.2), IL-4 (11B11), PD-1(RMP1-30), and ICOS(D10.G4.1) antibodies were purchased from BioLegend.Fluorescence-conjugated anti-mouse TCRβ (H57-597), NK1.1 (PK136), CD44(IM7), B220(RA3-6B2), Gr-1(RB6-8C5), CD11c(N418), CD11b(M1/70), CD4 (GK1.5), CD8 (53-6.7), IFN-γ (XMG1.2), IL-4 (11B11), PD-1(RMP1-30), and ICOS(D10.G4.1) antibodies were purchased from BioLegend.. Anti–phospho-p44/42 mitogen-activated protein kinase T202/Y204 (197G2), anti–phospho-S6 S240/244 (D68F8), anti–phospho-4E-BP1 T37/46 (236B4), anti–phospho-Akt S473 (193H12), TSC1 (D43E2), and TSC2 (D93F12) were purchased from Cell Signaling Technology, Inc. Anti–β-actin antibody was purchased from Sigma-Aldrich.. Cell surface staining was performed with 2% FBS-PBS.Cell surface staining was performed with 2% FBS-PBS.

    Article Title: TSC1 promotes i NKT cell anergy and inhibits i NKT cell-mediated anti-tumor immunity
    Article Snippet: CD11b(M1/70), CD4 (GK1.5), CD8 (53-6.7), IFN-γ (XMG1.2), IL-4 (11B11), PD-1(RMP1-30), and ICOS(D10.G4.1) antibodies were purchased from BioLegend. .. Anti–phospho-p44/42 mitogen-activated protein kinase T202/Y204 (197G2), anti–phospho-S6 S240/244 (D68F8), anti–phospho-4E-BP1 T37/46 (236B4), anti–phospho-Akt S473 (193H12), TSC1 (D43E2), and TSC2 (D93F12) were purchased from Cell Signaling Technology, Inc. Anti–β-actin antibody was purchased from Sigma-Aldrich. .. Cell surface staining was performed with 2% FBS-PBS.

    Quantitative RT-PCR:

    Article Title: Trans‐gnetin H isolated from the seeds of Paeonia species induces autophagy via inhibiting mTORC1 signalling through AMPK activation
    Article Snippet: The WB protocol was the same as our previous work.The WB protocol was the same as our previous work.. The primary antibodies Actin (20536‐1‐AP), LC3II (18725‐1‐AP), p62 (18420‐1‐AP) were purchased from Proteintech; TFEB (37785 S), pT389‐S6K (9234 S), p‐S6 (4858 S), S6K (9202 S), S6 (2217 S), p‐AMPKα (50081 S), AMPKα (5832 S), Raptor (2280 S) and TSC2 (D93F12) were purchased from Cell Signaling Technology; Flag (db7002) and HA (db2603) were purchased from Hangzhou Bio Technolog.. The secondary antibodies were obtained from Sigma.The secondary antibodies were obtained from Sigma.

    Article Title: A translational program that suppresses metabolism to shield the genome
    Article Snippet: Blots were stripped and reprobed when looking at total versus modified proteins unless otherwise noted.. Antibodies (all 1:1000): GLUT1 (Novus Biologicals, # NB110-39113), PAI-1 (Proteintech, #13801-1-AP), Puromycin (3RH11) (Kerafast, #EQ0001), β-ACTIN (C4) (SantaCruz Biotechnology, #sc-47778), EIF5A (abcam, #ab32443), EIF3D (Proteintech, # 10219-1-AP), EIF3K (Proteintech, # 10640-1-AP), acetyl-EIF5A (Lys47) (Boster Bio, #P01727), CYR61 (Proteintech, #26689-1-AP), SDC4 (R&D Systems, #P31431), SIRT1 (Proteintech, #13161-1-AP), SIRT2 (Proteintech, #19655-1-AP), NFKB2 (Proteintech, #10409-2-AP), 4E-BP (Cell Signaling, #9452 S), Phospho-4E-BP (Ser65) (Cell Signaling, #9451 S), TSC2 (D93F12) (Cell Signaling, #4308S), gamma H2AX (Novus, NB100-384), H2AX (Abcam, Ab11175), CRM-1 (Novus, NB100-79802), XPO-4 (Abcam, ab133237), eIF5A2 (Thermo, #PA5-30770), Phospho-RAPTOR (Ser792) (Cell Signaling, #89146S), RAPTOR (Cell Signaling, #2280), Phospho-Tuberin/TSC2 (Ser1387) (Cell Signaling, #5584S), NDRG1 (Abcam, #ab37897), and Anti-hypusine (mAbHpu24) antibody was kindly provided by Genentech.. The eIF5A1 antibodies and siRNAs used in this study were specific to eIF5A1 and not eIF5A2 as described by the manufacturer and validated in Supplementary Fig. .

    Article Title: Identification of mechanically regulated phosphorylation sites on tuberin (TSC2) that control mechanistic target of rapamycin (mTOR) signaling
    Article Snippet: Images were quantified using ImageJ software (U.S. National Institutes of Health, Bethesda, MD; http://rsb.info.nih.gov/nih-image/).Images were quantified using ImageJ software (U.S. National Institutes of Health, Bethesda, MD; http://rsb.info.nih.gov/nih-image/).. Antibodies - P-p70(389) (1A5) (#9206, used to probe Myc-tagged P-p70), P-p70(389) (#9234, used to probe endogenous P-p70), total p70 (49D7) (#2708), Anti-P-Akt substrate (RxRxxS*/T*) (23C8D2) for detection of PTSC2 (#10001), P-JNK(Thr183/Tyr185) (98F2) (#4671), total JNK (#9252), total mTOR (#2972), total Rictor (D16H9) (#9476), P-PKB (Ser473) (#4060), P-PKB (Thr308) (#9275S), total PKB (#9272), P-S6 (Ser240/4) (#5364), P-S6 (Ser235/6) (#2211), total S6 (#2217S), total TSC1 (D43E2) (#6935), total TSC2 (D93F12) (#4308) were purchased from Cell Signaling Technology (Danvers, MA, USA).. Anti-puromycin (12D10) (#MABE343) was purchased from EMD Millipore (Billerica, MA, USA).Anti-puromycin (12D10) (#MABE343) was purchased from EMD Millipore (Billerica, MA, USA).

    Article Title: Chronic signaling via the metabolic checkpoint kinase mTORC1 induces macrophage granuloma formation and marks sarcoidosis progression
    Article Snippet: HRP-conjugated secondary antibodies (Bethyl) 1:10000 were applied for 1 hour at room temperature in 4% low-fat milk and proteins were visualized using ECL substrate (Thermo Scientific).HRP-conjugated secondary antibodies (Bethyl) 1:10000 were applied for 1 hour at room temperature in 4% low-fat milk and proteins were visualized using ECL substrate (Thermo Scientific).. Primary antibodies were TSC2 (D93F12) 1:1000, p-Akt S473 (D9E) 1:1000, 4E-BP1 (53H11) 1:1000, p-S6 S240/244 (rabbit polyclonal) 1:3000, S6 (54D2) 1:1000, p-Akt T308 (D25E6) 1:1000, pan-Akt (40D4) 1:1000, β-tubulin (9F3) 1:1000 , p-RB Ser807/811 (D20B12) 1:1000, PDCD4 (D29C6) 1:1000, cleaved caspase 3 Asp175 (5A1E or rabbit polyclonal) 1:500, survivin (71G4B7) 1:1000 and p-S6K1 T389 (108D2) 1:1000, p-TSC2 S939 (rabbit polyclonal) 1:1000 obtained from Cell Signaling Technology.. Bcl-2 (N-19) 1:500, cyclin D1 (M-20) 1:500, p27Kip1 (C-19) 1:1000, cyclin A (C-19) 1:500, CDK4 (C-22) 1:1000, CDK6 (H-96) 1:500, and E2F1 (KH95) 1:500 were fom Santa Cruz. α-tubulin (CP06) 1:3000 was purchased from Calbiochem and GAPDH (rabbit polyclonal) 1:10000 from Trevigen.Bcl-2 (N-19) 1:500, cyclin D1 (M-20) 1:500, p27Kip1 (C-19) 1:1000, cyclin A (C-19) 1:500, CDK4 (C-22) 1:1000, CDK6 (H-96) 1:500, and E2F1 (KH95) 1:500 were fom Santa Cruz. α-tubulin (CP06) 1:3000 was purchased..

    Article Title: TSC1 promotes i NKT cell anergy and inhibits i NKT cell-mediated anti-tumor immunity
    Article Snippet: Fluorescence-conjugated anti-mouse TCRβ (H57-597), NK1.1 (PK136), CD44(IM7), B220(RA3-6B2), Gr-1(RB6-8C5), CD11c(N418), CD11b(M1/70), CD4 (GK1.5), CD8 (53-6.7), IFN-γ (XMG1.2), IL-4 (11B11), PD-1(RMP1-30), and ICOS(D10.G4.1) antibodies were purchased from BioLegend.Fluorescence-conjugated anti-mouse TCRβ (H57-597), NK1.1 (PK136), CD44(IM7), B220(RA3-6B2), Gr-1(RB6-8C5), CD11c(N418), CD11b(M1/70), CD4 (GK1.5), CD8 (53-6.7), IFN-γ (XMG1.2), IL-4 (11B11), PD-1(RMP1-30), and ICOS(D10.G4.1) antibodies were purchased from BioLegend.. Anti–phospho-p44/42 mitogen-activated protein kinase T202/Y204 (197G2), anti–phospho-S6 S240/244 (D68F8), anti–phospho-4E-BP1 T37/46 (236B4), anti–phospho-Akt S473 (193H12), TSC1 (D43E2), and TSC2 (D93F12) were purchased from Cell Signaling Technology, Inc. Anti–β-actin antibody was purchased from Sigma-Aldrich.. Cell surface staining was performed with 2% FBS-PBS.Cell surface staining was performed with 2% FBS-PBS.

    Article Title: TSC1 promotes i NKT cell anergy and inhibits i NKT cell-mediated anti-tumor immunity
    Article Snippet: CD11b(M1/70), CD4 (GK1.5), CD8 (53-6.7), IFN-γ (XMG1.2), IL-4 (11B11), PD-1(RMP1-30), and ICOS(D10.G4.1) antibodies were purchased from BioLegend. .. Anti–phospho-p44/42 mitogen-activated protein kinase T202/Y204 (197G2), anti–phospho-S6 S240/244 (D68F8), anti–phospho-4E-BP1 T37/46 (236B4), anti–phospho-Akt S473 (193H12), TSC1 (D43E2), and TSC2 (D93F12) were purchased from Cell Signaling Technology, Inc. Anti–β-actin antibody was purchased from Sigma-Aldrich. .. Cell surface staining was performed with 2% FBS-PBS.

    Western Blot:

    Article Title: Trans‐gnetin H isolated from the seeds of Paeonia species induces autophagy via inhibiting mTORC1 signalling through AMPK activation
    Article Snippet: The WB protocol was the same as our previous work.The WB protocol was the same as our previous work.. The primary antibodies Actin (20536‐1‐AP), LC3II (18725‐1‐AP), p62 (18420‐1‐AP) were purchased from Proteintech; TFEB (37785 S), pT389‐S6K (9234 S), p‐S6 (4858 S), S6K (9202 S), S6 (2217 S), p‐AMPKα (50081 S), AMPKα (5832 S), Raptor (2280 S) and TSC2 (D93F12) were purchased from Cell Signaling Technology; Flag (db7002) and HA (db2603) were purchased from Hangzhou Bio Technolog.. The secondary antibodies were obtained from Sigma.The secondary antibodies were obtained from Sigma.

    Article Title: A translational program that suppresses metabolism to shield the genome
    Article Snippet: Blots were stripped and reprobed when looking at total versus modified proteins unless otherwise noted.. Antibodies (all 1:1000): GLUT1 (Novus Biologicals, # NB110-39113), PAI-1 (Proteintech, #13801-1-AP), Puromycin (3RH11) (Kerafast, #EQ0001), β-ACTIN (C4) (SantaCruz Biotechnology, #sc-47778), EIF5A (abcam, #ab32443), EIF3D (Proteintech, # 10219-1-AP), EIF3K (Proteintech, # 10640-1-AP), acetyl-EIF5A (Lys47) (Boster Bio, #P01727), CYR61 (Proteintech, #26689-1-AP), SDC4 (R&D Systems, #P31431), SIRT1 (Proteintech, #13161-1-AP), SIRT2 (Proteintech, #19655-1-AP), NFKB2 (Proteintech, #10409-2-AP), 4E-BP (Cell Signaling, #9452 S), Phospho-4E-BP (Ser65) (Cell Signaling, #9451 S), TSC2 (D93F12) (Cell Signaling, #4308S), gamma H2AX (Novus, NB100-384), H2AX (Abcam, Ab11175), CRM-1 (Novus, NB100-79802), XPO-4 (Abcam, ab133237), eIF5A2 (Thermo, #PA5-30770), Phospho-RAPTOR (Ser792) (Cell Signaling, #89146S), RAPTOR (Cell Signaling, #2280), Phospho-Tuberin/TSC2 (Ser1387) (Cell Signaling, #5584S), NDRG1 (Abcam, #ab37897), and Anti-hypusine (mAbHpu24) antibody was kindly provided by Genentech.. The eIF5A1 antibodies and siRNAs used in this study were specific to eIF5A1 and not eIF5A2 as described by the manufacturer and validated in Supplementary Fig. .

    Article Title: Identification of mechanically regulated phosphorylation sites on tuberin (TSC2) that control mechanistic target of rapamycin (mTOR) signaling
    Article Snippet: Images were quantified using ImageJ software (U.S. National Institutes of Health, Bethesda, MD; http://rsb.info.nih.gov/nih-image/).Images were quantified using ImageJ software (U.S. National Institutes of Health, Bethesda, MD; http://rsb.info.nih.gov/nih-image/).. Antibodies - P-p70(389) (1A5) (#9206, used to probe Myc-tagged P-p70), P-p70(389) (#9234, used to probe endogenous P-p70), total p70 (49D7) (#2708), Anti-P-Akt substrate (RxRxxS*/T*) (23C8D2) for detection of PTSC2 (#10001), P-JNK(Thr183/Tyr185) (98F2) (#4671), total JNK (#9252), total mTOR (#2972), total Rictor (D16H9) (#9476), P-PKB (Ser473) (#4060), P-PKB (Thr308) (#9275S), total PKB (#9272), P-S6 (Ser240/4) (#5364), P-S6 (Ser235/6) (#2211), total S6 (#2217S), total TSC1 (D43E2) (#6935), total TSC2 (D93F12) (#4308) were purchased from Cell Signaling Technology (Danvers, MA, USA).. Anti-puromycin (12D10) (#MABE343) was purchased from EMD Millipore (Billerica, MA, USA).Anti-puromycin (12D10) (#MABE343) was purchased from EMD Millipore (Billerica, MA, USA).

    Article Title: Chronic signaling via the metabolic checkpoint kinase mTORC1 induces macrophage granuloma formation and marks sarcoidosis progression
    Article Snippet: HRP-conjugated secondary antibodies (Bethyl) 1:10000 were applied for 1 hour at room temperature in 4% low-fat milk and proteins were visualized using ECL substrate (Thermo Scientific).HRP-conjugated secondary antibodies (Bethyl) 1:10000 were applied for 1 hour at room temperature in 4% low-fat milk and proteins were visualized using ECL substrate (Thermo Scientific).. Primary antibodies were TSC2 (D93F12) 1:1000, p-Akt S473 (D9E) 1:1000, 4E-BP1 (53H11) 1:1000, p-S6 S240/244 (rabbit polyclonal) 1:3000, S6 (54D2) 1:1000, p-Akt T308 (D25E6) 1:1000, pan-Akt (40D4) 1:1000, β-tubulin (9F3) 1:1000 , p-RB Ser807/811 (D20B12) 1:1000, PDCD4 (D29C6) 1:1000, cleaved caspase 3 Asp175 (5A1E or rabbit polyclonal) 1:500, survivin (71G4B7) 1:1000 and p-S6K1 T389 (108D2) 1:1000, p-TSC2 S939 (rabbit polyclonal) 1:1000 obtained from Cell Signaling Technology.. Bcl-2 (N-19) 1:500, cyclin D1 (M-20) 1:500, p27Kip1 (C-19) 1:1000, cyclin A (C-19) 1:500, CDK4 (C-22) 1:1000, CDK6 (H-96) 1:500, and E2F1 (KH95) 1:500 were fom Santa Cruz. α-tubulin (CP06) 1:3000 was purchased from Calbiochem and GAPDH (rabbit polyclonal) 1:10000 from Trevigen.Bcl-2 (N-19) 1:500, cyclin D1 (M-20) 1:500, p27Kip1 (C-19) 1:1000, cyclin A (C-19) 1:500, CDK4 (C-22) 1:1000, CDK6 (H-96) 1:500, and E2F1 (KH95) 1:500 were fom Santa Cruz. α-tubulin (CP06) 1:3000 was purchased..

    Article Title: TSC1 promotes i NKT cell anergy and inhibits i NKT cell-mediated anti-tumor immunity
    Article Snippet: Fluorescence-conjugated anti-mouse TCRβ (H57-597), NK1.1 (PK136), CD44(IM7), B220(RA3-6B2), Gr-1(RB6-8C5), CD11c(N418), CD11b(M1/70), CD4 (GK1.5), CD8 (53-6.7), IFN-γ (XMG1.2), IL-4 (11B11), PD-1(RMP1-30), and ICOS(D10.G4.1) antibodies were purchased from BioLegend.Fluorescence-conjugated anti-mouse TCRβ (H57-597), NK1.1 (PK136), CD44(IM7), B220(RA3-6B2), Gr-1(RB6-8C5), CD11c(N418), CD11b(M1/70), CD4 (GK1.5), CD8 (53-6.7), IFN-γ (XMG1.2), IL-4 (11B11), PD-1(RMP1-30), and ICOS(D10.G4.1) antibodies were purchased from BioLegend.. Anti–phospho-p44/42 mitogen-activated protein kinase T202/Y204 (197G2), anti–phospho-S6 S240/244 (D68F8), anti–phospho-4E-BP1 T37/46 (236B4), anti–phospho-Akt S473 (193H12), TSC1 (D43E2), and TSC2 (D93F12) were purchased from Cell Signaling Technology, Inc. Anti–β-actin antibody was purchased from Sigma-Aldrich.. Cell surface staining was performed with 2% FBS-PBS.Cell surface staining was performed with 2% FBS-PBS.

    Article Title: TSC1 promotes i NKT cell anergy and inhibits i NKT cell-mediated anti-tumor immunity
    Article Snippet: CD11b(M1/70), CD4 (GK1.5), CD8 (53-6.7), IFN-γ (XMG1.2), IL-4 (11B11), PD-1(RMP1-30), and ICOS(D10.G4.1) antibodies were purchased from BioLegend. .. Anti–phospho-p44/42 mitogen-activated protein kinase T202/Y204 (197G2), anti–phospho-S6 S240/244 (D68F8), anti–phospho-4E-BP1 T37/46 (236B4), anti–phospho-Akt S473 (193H12), TSC1 (D43E2), and TSC2 (D93F12) were purchased from Cell Signaling Technology, Inc. Anti–β-actin antibody was purchased from Sigma-Aldrich. .. Cell surface staining was performed with 2% FBS-PBS.

    Immunocytochemistry:

    Article Title: Trans‐gnetin H isolated from the seeds of Paeonia species induces autophagy via inhibiting mTORC1 signalling through AMPK activation
    Article Snippet: The WB protocol was the same as our previous work.The WB protocol was the same as our previous work.. The primary antibodies Actin (20536‐1‐AP), LC3II (18725‐1‐AP), p62 (18420‐1‐AP) were purchased from Proteintech; TFEB (37785 S), pT389‐S6K (9234 S), p‐S6 (4858 S), S6K (9202 S), S6 (2217 S), p‐AMPKα (50081 S), AMPKα (5832 S), Raptor (2280 S) and TSC2 (D93F12) were purchased from Cell Signaling Technology; Flag (db7002) and HA (db2603) were purchased from Hangzhou Bio Technolog.. The secondary antibodies were obtained from Sigma.The secondary antibodies were obtained from Sigma.

    Article Title: A translational program that suppresses metabolism to shield the genome
    Article Snippet: Blots were stripped and reprobed when looking at total versus modified proteins unless otherwise noted.. Antibodies (all 1:1000): GLUT1 (Novus Biologicals, # NB110-39113), PAI-1 (Proteintech, #13801-1-AP), Puromycin (3RH11) (Kerafast, #EQ0001), β-ACTIN (C4) (SantaCruz Biotechnology, #sc-47778), EIF5A (abcam, #ab32443), EIF3D (Proteintech, # 10219-1-AP), EIF3K (Proteintech, # 10640-1-AP), acetyl-EIF5A (Lys47) (Boster Bio, #P01727), CYR61 (Proteintech, #26689-1-AP), SDC4 (R&D Systems, #P31431), SIRT1 (Proteintech, #13161-1-AP), SIRT2 (Proteintech, #19655-1-AP), NFKB2 (Proteintech, #10409-2-AP), 4E-BP (Cell Signaling, #9452 S), Phospho-4E-BP (Ser65) (Cell Signaling, #9451 S), TSC2 (D93F12) (Cell Signaling, #4308S), gamma H2AX (Novus, NB100-384), H2AX (Abcam, Ab11175), CRM-1 (Novus, NB100-79802), XPO-4 (Abcam, ab133237), eIF5A2 (Thermo, #PA5-30770), Phospho-RAPTOR (Ser792) (Cell Signaling, #89146S), RAPTOR (Cell Signaling, #2280), Phospho-Tuberin/TSC2 (Ser1387) (Cell Signaling, #5584S), NDRG1 (Abcam, #ab37897), and Anti-hypusine (mAbHpu24) antibody was kindly provided by Genentech.. The eIF5A1 antibodies and siRNAs used in this study were specific to eIF5A1 and not eIF5A2 as described by the manufacturer and validated in Supplementary Fig. .

    Article Title: Identification of mechanically regulated phosphorylation sites on tuberin (TSC2) that control mechanistic target of rapamycin (mTOR) signaling
    Article Snippet: Images were quantified using ImageJ software (U.S. National Institutes of Health, Bethesda, MD; http://rsb.info.nih.gov/nih-image/).Images were quantified using ImageJ software (U.S. National Institutes of Health, Bethesda, MD; http://rsb.info.nih.gov/nih-image/).. Antibodies - P-p70(389) (1A5) (#9206, used to probe Myc-tagged P-p70), P-p70(389) (#9234, used to probe endogenous P-p70), total p70 (49D7) (#2708), Anti-P-Akt substrate (RxRxxS*/T*) (23C8D2) for detection of PTSC2 (#10001), P-JNK(Thr183/Tyr185) (98F2) (#4671), total JNK (#9252), total mTOR (#2972), total Rictor (D16H9) (#9476), P-PKB (Ser473) (#4060), P-PKB (Thr308) (#9275S), total PKB (#9272), P-S6 (Ser240/4) (#5364), P-S6 (Ser235/6) (#2211), total S6 (#2217S), total TSC1 (D43E2) (#6935), total TSC2 (D93F12) (#4308) were purchased from Cell Signaling Technology (Danvers, MA, USA).. Anti-puromycin (12D10) (#MABE343) was purchased from EMD Millipore (Billerica, MA, USA).Anti-puromycin (12D10) (#MABE343) was purchased from EMD Millipore (Billerica, MA, USA).

    Article Title: Chronic signaling via the metabolic checkpoint kinase mTORC1 induces macrophage granuloma formation and marks sarcoidosis progression
    Article Snippet: HRP-conjugated secondary antibodies (Bethyl) 1:10000 were applied for 1 hour at room temperature in 4% low-fat milk and proteins were visualized using ECL substrate (Thermo Scientific).HRP-conjugated secondary antibodies (Bethyl) 1:10000 were applied for 1 hour at room temperature in 4% low-fat milk and proteins were visualized using ECL substrate (Thermo Scientific).. Primary antibodies were TSC2 (D93F12) 1:1000, p-Akt S473 (D9E) 1:1000, 4E-BP1 (53H11) 1:1000, p-S6 S240/244 (rabbit polyclonal) 1:3000, S6 (54D2) 1:1000, p-Akt T308 (D25E6) 1:1000, pan-Akt (40D4) 1:1000, β-tubulin (9F3) 1:1000 , p-RB Ser807/811 (D20B12) 1:1000, PDCD4 (D29C6) 1:1000, cleaved caspase 3 Asp175 (5A1E or rabbit polyclonal) 1:500, survivin (71G4B7) 1:1000 and p-S6K1 T389 (108D2) 1:1000, p-TSC2 S939 (rabbit polyclonal) 1:1000 obtained from Cell Signaling Technology.. Bcl-2 (N-19) 1:500, cyclin D1 (M-20) 1:500, p27Kip1 (C-19) 1:1000, cyclin A (C-19) 1:500, CDK4 (C-22) 1:1000, CDK6 (H-96) 1:500, and E2F1 (KH95) 1:500 were fom Santa Cruz. α-tubulin (CP06) 1:3000 was purchased from Calbiochem and GAPDH (rabbit polyclonal) 1:10000 from Trevigen.Bcl-2 (N-19) 1:500, cyclin D1 (M-20) 1:500, p27Kip1 (C-19) 1:1000, cyclin A (C-19) 1:500, CDK4 (C-22) 1:1000, CDK6 (H-96) 1:500, and E2F1 (KH95) 1:500 were fom Santa Cruz. α-tubulin (CP06) 1:3000 was purchased..

    Article Title: TSC1 promotes i NKT cell anergy and inhibits i NKT cell-mediated anti-tumor immunity
    Article Snippet: Fluorescence-conjugated anti-mouse TCRβ (H57-597), NK1.1 (PK136), CD44(IM7), B220(RA3-6B2), Gr-1(RB6-8C5), CD11c(N418), CD11b(M1/70), CD4 (GK1.5), CD8 (53-6.7), IFN-γ (XMG1.2), IL-4 (11B11), PD-1(RMP1-30), and ICOS(D10.G4.1) antibodies were purchased from BioLegend.Fluorescence-conjugated anti-mouse TCRβ (H57-597), NK1.1 (PK136), CD44(IM7), B220(RA3-6B2), Gr-1(RB6-8C5), CD11c(N418), CD11b(M1/70), CD4 (GK1.5), CD8 (53-6.7), IFN-γ (XMG1.2), IL-4 (11B11), PD-1(RMP1-30), and ICOS(D10.G4.1) antibodies were purchased from BioLegend.. Anti–phospho-p44/42 mitogen-activated protein kinase T202/Y204 (197G2), anti–phospho-S6 S240/244 (D68F8), anti–phospho-4E-BP1 T37/46 (236B4), anti–phospho-Akt S473 (193H12), TSC1 (D43E2), and TSC2 (D93F12) were purchased from Cell Signaling Technology, Inc. Anti–β-actin antibody was purchased from Sigma-Aldrich.. Cell surface staining was performed with 2% FBS-PBS.Cell surface staining was performed with 2% FBS-PBS.

    Article Title: TSC1 promotes i NKT cell anergy and inhibits i NKT cell-mediated anti-tumor immunity
    Article Snippet: CD11b(M1/70), CD4 (GK1.5), CD8 (53-6.7), IFN-γ (XMG1.2), IL-4 (11B11), PD-1(RMP1-30), and ICOS(D10.G4.1) antibodies were purchased from BioLegend. .. Anti–phospho-p44/42 mitogen-activated protein kinase T202/Y204 (197G2), anti–phospho-S6 S240/244 (D68F8), anti–phospho-4E-BP1 T37/46 (236B4), anti–phospho-Akt S473 (193H12), TSC1 (D43E2), and TSC2 (D93F12) were purchased from Cell Signaling Technology, Inc. Anti–β-actin antibody was purchased from Sigma-Aldrich. .. Cell surface staining was performed with 2% FBS-PBS.

    Staining:

    Article Title: Trans‐gnetin H isolated from the seeds of Paeonia species induces autophagy via inhibiting mTORC1 signalling through AMPK activation
    Article Snippet: The WB protocol was the same as our previous work.The WB protocol was the same as our previous work.. The primary antibodies Actin (20536‐1‐AP), LC3II (18725‐1‐AP), p62 (18420‐1‐AP) were purchased from Proteintech; TFEB (37785 S), pT389‐S6K (9234 S), p‐S6 (4858 S), S6K (9202 S), S6 (2217 S), p‐AMPKα (50081 S), AMPKα (5832 S), Raptor (2280 S) and TSC2 (D93F12) were purchased from Cell Signaling Technology; Flag (db7002) and HA (db2603) were purchased from Hangzhou Bio Technolog.. The secondary antibodies were obtained from Sigma.The secondary antibodies were obtained from Sigma.

    Article Title: A translational program that suppresses metabolism to shield the genome
    Article Snippet: Blots were stripped and reprobed when looking at total versus modified proteins unless otherwise noted.. Antibodies (all 1:1000): GLUT1 (Novus Biologicals, # NB110-39113), PAI-1 (Proteintech, #13801-1-AP), Puromycin (3RH11) (Kerafast, #EQ0001), β-ACTIN (C4) (SantaCruz Biotechnology, #sc-47778), EIF5A (abcam, #ab32443), EIF3D (Proteintech, # 10219-1-AP), EIF3K (Proteintech, # 10640-1-AP), acetyl-EIF5A (Lys47) (Boster Bio, #P01727), CYR61 (Proteintech, #26689-1-AP), SDC4 (R&D Systems, #P31431), SIRT1 (Proteintech, #13161-1-AP), SIRT2 (Proteintech, #19655-1-AP), NFKB2 (Proteintech, #10409-2-AP), 4E-BP (Cell Signaling, #9452 S), Phospho-4E-BP (Ser65) (Cell Signaling, #9451 S), TSC2 (D93F12) (Cell Signaling, #4308S), gamma H2AX (Novus, NB100-384), H2AX (Abcam, Ab11175), CRM-1 (Novus, NB100-79802), XPO-4 (Abcam, ab133237), eIF5A2 (Thermo, #PA5-30770), Phospho-RAPTOR (Ser792) (Cell Signaling, #89146S), RAPTOR (Cell Signaling, #2280), Phospho-Tuberin/TSC2 (Ser1387) (Cell Signaling, #5584S), NDRG1 (Abcam, #ab37897), and Anti-hypusine (mAbHpu24) antibody was kindly provided by Genentech.. The eIF5A1 antibodies and siRNAs used in this study were specific to eIF5A1 and not eIF5A2 as described by the manufacturer and validated in Supplementary Fig. .

    Article Title: Identification of mechanically regulated phosphorylation sites on tuberin (TSC2) that control mechanistic target of rapamycin (mTOR) signaling
    Article Snippet: Images were quantified using ImageJ software (U.S. National Institutes of Health, Bethesda, MD; http://rsb.info.nih.gov/nih-image/).Images were quantified using ImageJ software (U.S. National Institutes of Health, Bethesda, MD; http://rsb.info.nih.gov/nih-image/).. Antibodies - P-p70(389) (1A5) (#9206, used to probe Myc-tagged P-p70), P-p70(389) (#9234, used to probe endogenous P-p70), total p70 (49D7) (#2708), Anti-P-Akt substrate (RxRxxS*/T*) (23C8D2) for detection of PTSC2 (#10001), P-JNK(Thr183/Tyr185) (98F2) (#4671), total JNK (#9252), total mTOR (#2972), total Rictor (D16H9) (#9476), P-PKB (Ser473) (#4060), P-PKB (Thr308) (#9275S), total PKB (#9272), P-S6 (Ser240/4) (#5364), P-S6 (Ser235/6) (#2211), total S6 (#2217S), total TSC1 (D43E2) (#6935), total TSC2 (D93F12) (#4308) were purchased from Cell Signaling Technology (Danvers, MA, USA).. Anti-puromycin (12D10) (#MABE343) was purchased from EMD Millipore (Billerica, MA, USA).Anti-puromycin (12D10) (#MABE343) was purchased from EMD Millipore (Billerica, MA, USA).

    Article Title: Chronic signaling via the metabolic checkpoint kinase mTORC1 induces macrophage granuloma formation and marks sarcoidosis progression
    Article Snippet: HRP-conjugated secondary antibodies (Bethyl) 1:10000 were applied for 1 hour at room temperature in 4% low-fat milk and proteins were visualized using ECL substrate (Thermo Scientific).HRP-conjugated secondary antibodies (Bethyl) 1:10000 were applied for 1 hour at room temperature in 4% low-fat milk and proteins were visualized using ECL substrate (Thermo Scientific).. Primary antibodies were TSC2 (D93F12) 1:1000, p-Akt S473 (D9E) 1:1000, 4E-BP1 (53H11) 1:1000, p-S6 S240/244 (rabbit polyclonal) 1:3000, S6 (54D2) 1:1000, p-Akt T308 (D25E6) 1:1000, pan-Akt (40D4) 1:1000, β-tubulin (9F3) 1:1000 , p-RB Ser807/811 (D20B12) 1:1000, PDCD4 (D29C6) 1:1000, cleaved caspase 3 Asp175 (5A1E or rabbit polyclonal) 1:500, survivin (71G4B7) 1:1000 and p-S6K1 T389 (108D2) 1:1000, p-TSC2 S939 (rabbit polyclonal) 1:1000 obtained from Cell Signaling Technology.. Bcl-2 (N-19) 1:500, cyclin D1 (M-20) 1:500, p27Kip1 (C-19) 1:1000, cyclin A (C-19) 1:500, CDK4 (C-22) 1:1000, CDK6 (H-96) 1:500, and E2F1 (KH95) 1:500 were fom Santa Cruz. α-tubulin (CP06) 1:3000 was purchased from Calbiochem and GAPDH (rabbit polyclonal) 1:10000 from Trevigen.Bcl-2 (N-19) 1:500, cyclin D1 (M-20) 1:500, p27Kip1 (C-19) 1:1000, cyclin A (C-19) 1:500, CDK4 (C-22) 1:1000, CDK6 (H-96) 1:500, and E2F1 (KH95) 1:500 were fom Santa Cruz. α-tubulin (CP06) 1:3000 was purchased..

    Article Title: TSC1 promotes i NKT cell anergy and inhibits i NKT cell-mediated anti-tumor immunity
    Article Snippet: Fluorescence-conjugated anti-mouse TCRβ (H57-597), NK1.1 (PK136), CD44(IM7), B220(RA3-6B2), Gr-1(RB6-8C5), CD11c(N418), CD11b(M1/70), CD4 (GK1.5), CD8 (53-6.7), IFN-γ (XMG1.2), IL-4 (11B11), PD-1(RMP1-30), and ICOS(D10.G4.1) antibodies were purchased from BioLegend.Fluorescence-conjugated anti-mouse TCRβ (H57-597), NK1.1 (PK136), CD44(IM7), B220(RA3-6B2), Gr-1(RB6-8C5), CD11c(N418), CD11b(M1/70), CD4 (GK1.5), CD8 (53-6.7), IFN-γ (XMG1.2), IL-4 (11B11), PD-1(RMP1-30), and ICOS(D10.G4.1) antibodies were purchased from BioLegend.. Anti–phospho-p44/42 mitogen-activated protein kinase T202/Y204 (197G2), anti–phospho-S6 S240/244 (D68F8), anti–phospho-4E-BP1 T37/46 (236B4), anti–phospho-Akt S473 (193H12), TSC1 (D43E2), and TSC2 (D93F12) were purchased from Cell Signaling Technology, Inc. Anti–β-actin antibody was purchased from Sigma-Aldrich.. Cell surface staining was performed with 2% FBS-PBS.Cell surface staining was performed with 2% FBS-PBS.

    Article Title: TSC1 promotes i NKT cell anergy and inhibits i NKT cell-mediated anti-tumor immunity
    Article Snippet: CD11b(M1/70), CD4 (GK1.5), CD8 (53-6.7), IFN-γ (XMG1.2), IL-4 (11B11), PD-1(RMP1-30), and ICOS(D10.G4.1) antibodies were purchased from BioLegend. .. Anti–phospho-p44/42 mitogen-activated protein kinase T202/Y204 (197G2), anti–phospho-S6 S240/244 (D68F8), anti–phospho-4E-BP1 T37/46 (236B4), anti–phospho-Akt S473 (193H12), TSC1 (D43E2), and TSC2 (D93F12) were purchased from Cell Signaling Technology, Inc. Anti–β-actin antibody was purchased from Sigma-Aldrich. .. Cell surface staining was performed with 2% FBS-PBS.

    Inhibition:

    Article Title: Trans‐gnetin H isolated from the seeds of Paeonia species induces autophagy via inhibiting mTORC1 signalling through AMPK activation
    Article Snippet: The WB protocol was the same as our previous work.The WB protocol was the same as our previous work.. The primary antibodies Actin (20536‐1‐AP), LC3II (18725‐1‐AP), p62 (18420‐1‐AP) were purchased from Proteintech; TFEB (37785 S), pT389‐S6K (9234 S), p‐S6 (4858 S), S6K (9202 S), S6 (2217 S), p‐AMPKα (50081 S), AMPKα (5832 S), Raptor (2280 S) and TSC2 (D93F12) were purchased from Cell Signaling Technology; Flag (db7002) and HA (db2603) were purchased from Hangzhou Bio Technolog.. The secondary antibodies were obtained from Sigma.The secondary antibodies were obtained from Sigma.

    Article Title: A translational program that suppresses metabolism to shield the genome
    Article Snippet: Blots were stripped and reprobed when looking at total versus modified proteins unless otherwise noted.. Antibodies (all 1:1000): GLUT1 (Novus Biologicals, # NB110-39113), PAI-1 (Proteintech, #13801-1-AP), Puromycin (3RH11) (Kerafast, #EQ0001), β-ACTIN (C4) (SantaCruz Biotechnology, #sc-47778), EIF5A (abcam, #ab32443), EIF3D (Proteintech, # 10219-1-AP), EIF3K (Proteintech, # 10640-1-AP), acetyl-EIF5A (Lys47) (Boster Bio, #P01727), CYR61 (Proteintech, #26689-1-AP), SDC4 (R&D Systems, #P31431), SIRT1 (Proteintech, #13161-1-AP), SIRT2 (Proteintech, #19655-1-AP), NFKB2 (Proteintech, #10409-2-AP), 4E-BP (Cell Signaling, #9452 S), Phospho-4E-BP (Ser65) (Cell Signaling, #9451 S), TSC2 (D93F12) (Cell Signaling, #4308S), gamma H2AX (Novus, NB100-384), H2AX (Abcam, Ab11175), CRM-1 (Novus, NB100-79802), XPO-4 (Abcam, ab133237), eIF5A2 (Thermo, #PA5-30770), Phospho-RAPTOR (Ser792) (Cell Signaling, #89146S), RAPTOR (Cell Signaling, #2280), Phospho-Tuberin/TSC2 (Ser1387) (Cell Signaling, #5584S), NDRG1 (Abcam, #ab37897), and Anti-hypusine (mAbHpu24) antibody was kindly provided by Genentech.. The eIF5A1 antibodies and siRNAs used in this study were specific to eIF5A1 and not eIF5A2 as described by the manufacturer and validated in Supplementary Fig. .

    Article Title: Identification of mechanically regulated phosphorylation sites on tuberin (TSC2) that control mechanistic target of rapamycin (mTOR) signaling
    Article Snippet: Images were quantified using ImageJ software (U.S. National Institutes of Health, Bethesda, MD; http://rsb.info.nih.gov/nih-image/).Images were quantified using ImageJ software (U.S. National Institutes of Health, Bethesda, MD; http://rsb.info.nih.gov/nih-image/).. Antibodies - P-p70(389) (1A5) (#9206, used to probe Myc-tagged P-p70), P-p70(389) (#9234, used to probe endogenous P-p70), total p70 (49D7) (#2708), Anti-P-Akt substrate (RxRxxS*/T*) (23C8D2) for detection of PTSC2 (#10001), P-JNK(Thr183/Tyr185) (98F2) (#4671), total JNK (#9252), total mTOR (#2972), total Rictor (D16H9) (#9476), P-PKB (Ser473) (#4060), P-PKB (Thr308) (#9275S), total PKB (#9272), P-S6 (Ser240/4) (#5364), P-S6 (Ser235/6) (#2211), total S6 (#2217S), total TSC1 (D43E2) (#6935), total TSC2 (D93F12) (#4308) were purchased from Cell Signaling Technology (Danvers, MA, USA).. Anti-puromycin (12D10) (#MABE343) was purchased from EMD Millipore (Billerica, MA, USA).Anti-puromycin (12D10) (#MABE343) was purchased from EMD Millipore (Billerica, MA, USA).

    Article Title: Chronic signaling via the metabolic checkpoint kinase mTORC1 induces macrophage granuloma formation and marks sarcoidosis progression
    Article Snippet: HRP-conjugated secondary antibodies (Bethyl) 1:10000 were applied for 1 hour at room temperature in 4% low-fat milk and proteins were visualized using ECL substrate (Thermo Scientific).HRP-conjugated secondary antibodies (Bethyl) 1:10000 were applied for 1 hour at room temperature in 4% low-fat milk and proteins were visualized using ECL substrate (Thermo Scientific).. Primary antibodies were TSC2 (D93F12) 1:1000, p-Akt S473 (D9E) 1:1000, 4E-BP1 (53H11) 1:1000, p-S6 S240/244 (rabbit polyclonal) 1:3000, S6 (54D2) 1:1000, p-Akt T308 (D25E6) 1:1000, pan-Akt (40D4) 1:1000, β-tubulin (9F3) 1:1000 , p-RB Ser807/811 (D20B12) 1:1000, PDCD4 (D29C6) 1:1000, cleaved caspase 3 Asp175 (5A1E or rabbit polyclonal) 1:500, survivin (71G4B7) 1:1000 and p-S6K1 T389 (108D2) 1:1000, p-TSC2 S939 (rabbit polyclonal) 1:1000 obtained from Cell Signaling Technology.. Bcl-2 (N-19) 1:500, cyclin D1 (M-20) 1:500, p27Kip1 (C-19) 1:1000, cyclin A (C-19) 1:500, CDK4 (C-22) 1:1000, CDK6 (H-96) 1:500, and E2F1 (KH95) 1:500 were fom Santa Cruz. α-tubulin (CP06) 1:3000 was purchased from Calbiochem and GAPDH (rabbit polyclonal) 1:10000 from Trevigen.Bcl-2 (N-19) 1:500, cyclin D1 (M-20) 1:500, p27Kip1 (C-19) 1:1000, cyclin A (C-19) 1:500, CDK4 (C-22) 1:1000, CDK6 (H-96) 1:500, and E2F1 (KH95) 1:500 were fom Santa Cruz. α-tubulin (CP06) 1:3000 was purchased..

    Article Title: TSC1 promotes i NKT cell anergy and inhibits i NKT cell-mediated anti-tumor immunity
    Article Snippet: Fluorescence-conjugated anti-mouse TCRβ (H57-597), NK1.1 (PK136), CD44(IM7), B220(RA3-6B2), Gr-1(RB6-8C5), CD11c(N418), CD11b(M1/70), CD4 (GK1.5), CD8 (53-6.7), IFN-γ (XMG1.2), IL-4 (11B11), PD-1(RMP1-30), and ICOS(D10.G4.1) antibodies were purchased from BioLegend.Fluorescence-conjugated anti-mouse TCRβ (H57-597), NK1.1 (PK136), CD44(IM7), B220(RA3-6B2), Gr-1(RB6-8C5), CD11c(N418), CD11b(M1/70), CD4 (GK1.5), CD8 (53-6.7), IFN-γ (XMG1.2), IL-4 (11B11), PD-1(RMP1-30), and ICOS(D10.G4.1) antibodies were purchased from BioLegend.. Anti–phospho-p44/42 mitogen-activated protein kinase T202/Y204 (197G2), anti–phospho-S6 S240/244 (D68F8), anti–phospho-4E-BP1 T37/46 (236B4), anti–phospho-Akt S473 (193H12), TSC1 (D43E2), and TSC2 (D93F12) were purchased from Cell Signaling Technology, Inc. Anti–β-actin antibody was purchased from Sigma-Aldrich.. Cell surface staining was performed with 2% FBS-PBS.Cell surface staining was performed with 2% FBS-PBS.

    Article Title: TSC1 promotes i NKT cell anergy and inhibits i NKT cell-mediated anti-tumor immunity
    Article Snippet: CD11b(M1/70), CD4 (GK1.5), CD8 (53-6.7), IFN-γ (XMG1.2), IL-4 (11B11), PD-1(RMP1-30), and ICOS(D10.G4.1) antibodies were purchased from BioLegend. .. Anti–phospho-p44/42 mitogen-activated protein kinase T202/Y204 (197G2), anti–phospho-S6 S240/244 (D68F8), anti–phospho-4E-BP1 T37/46 (236B4), anti–phospho-Akt S473 (193H12), TSC1 (D43E2), and TSC2 (D93F12) were purchased from Cell Signaling Technology, Inc. Anti–β-actin antibody was purchased from Sigma-Aldrich. .. Cell surface staining was performed with 2% FBS-PBS.

    TUNEL Assay:

    Article Title: Trans‐gnetin H isolated from the seeds of Paeonia species induces autophagy via inhibiting mTORC1 signalling through AMPK activation
    Article Snippet: The WB protocol was the same as our previous work.The WB protocol was the same as our previous work.. The primary antibodies Actin (20536‐1‐AP), LC3II (18725‐1‐AP), p62 (18420‐1‐AP) were purchased from Proteintech; TFEB (37785 S), pT389‐S6K (9234 S), p‐S6 (4858 S), S6K (9202 S), S6 (2217 S), p‐AMPKα (50081 S), AMPKα (5832 S), Raptor (2280 S) and TSC2 (D93F12) were purchased from Cell Signaling Technology; Flag (db7002) and HA (db2603) were purchased from Hangzhou Bio Technolog.. The secondary antibodies were obtained from Sigma.The secondary antibodies were obtained from Sigma.

    Article Title: A translational program that suppresses metabolism to shield the genome
    Article Snippet: Blots were stripped and reprobed when looking at total versus modified proteins unless otherwise noted.. Antibodies (all 1:1000): GLUT1 (Novus Biologicals, # NB110-39113), PAI-1 (Proteintech, #13801-1-AP), Puromycin (3RH11) (Kerafast, #EQ0001), β-ACTIN (C4) (SantaCruz Biotechnology, #sc-47778), EIF5A (abcam, #ab32443), EIF3D (Proteintech, # 10219-1-AP), EIF3K (Proteintech, # 10640-1-AP), acetyl-EIF5A (Lys47) (Boster Bio, #P01727), CYR61 (Proteintech, #26689-1-AP), SDC4 (R&D Systems, #P31431), SIRT1 (Proteintech, #13161-1-AP), SIRT2 (Proteintech, #19655-1-AP), NFKB2 (Proteintech, #10409-2-AP), 4E-BP (Cell Signaling, #9452 S), Phospho-4E-BP (Ser65) (Cell Signaling, #9451 S), TSC2 (D93F12) (Cell Signaling, #4308S), gamma H2AX (Novus, NB100-384), H2AX (Abcam, Ab11175), CRM-1 (Novus, NB100-79802), XPO-4 (Abcam, ab133237), eIF5A2 (Thermo, #PA5-30770), Phospho-RAPTOR (Ser792) (Cell Signaling, #89146S), RAPTOR (Cell Signaling, #2280), Phospho-Tuberin/TSC2 (Ser1387) (Cell Signaling, #5584S), NDRG1 (Abcam, #ab37897), and Anti-hypusine (mAbHpu24) antibody was kindly provided by Genentech.. The eIF5A1 antibodies and siRNAs used in this study were specific to eIF5A1 and not eIF5A2 as described by the manufacturer and validated in Supplementary Fig. .

    Article Title: Identification of mechanically regulated phosphorylation sites on tuberin (TSC2) that control mechanistic target of rapamycin (mTOR) signaling
    Article Snippet: Images were quantified using ImageJ software (U.S. National Institutes of Health, Bethesda, MD; http://rsb.info.nih.gov/nih-image/).Images were quantified using ImageJ software (U.S. National Institutes of Health, Bethesda, MD; http://rsb.info.nih.gov/nih-image/).. Antibodies - P-p70(389) (1A5) (#9206, used to probe Myc-tagged P-p70), P-p70(389) (#9234, used to probe endogenous P-p70), total p70 (49D7) (#2708), Anti-P-Akt substrate (RxRxxS*/T*) (23C8D2) for detection of PTSC2 (#10001), P-JNK(Thr183/Tyr185) (98F2) (#4671), total JNK (#9252), total mTOR (#2972), total Rictor (D16H9) (#9476), P-PKB (Ser473) (#4060), P-PKB (Thr308) (#9275S), total PKB (#9272), P-S6 (Ser240/4) (#5364), P-S6 (Ser235/6) (#2211), total S6 (#2217S), total TSC1 (D43E2) (#6935), total TSC2 (D93F12) (#4308) were purchased from Cell Signaling Technology (Danvers, MA, USA).. Anti-puromycin (12D10) (#MABE343) was purchased from EMD Millipore (Billerica, MA, USA).Anti-puromycin (12D10) (#MABE343) was purchased from EMD Millipore (Billerica, MA, USA).

    Article Title: Chronic signaling via the metabolic checkpoint kinase mTORC1 induces macrophage granuloma formation and marks sarcoidosis progression
    Article Snippet: HRP-conjugated secondary antibodies (Bethyl) 1:10000 were applied for 1 hour at room temperature in 4% low-fat milk and proteins were visualized using ECL substrate (Thermo Scientific).HRP-conjugated secondary antibodies (Bethyl) 1:10000 were applied for 1 hour at room temperature in 4% low-fat milk and proteins were visualized using ECL substrate (Thermo Scientific).. Primary antibodies were TSC2 (D93F12) 1:1000, p-Akt S473 (D9E) 1:1000, 4E-BP1 (53H11) 1:1000, p-S6 S240/244 (rabbit polyclonal) 1:3000, S6 (54D2) 1:1000, p-Akt T308 (D25E6) 1:1000, pan-Akt (40D4) 1:1000, β-tubulin (9F3) 1:1000 , p-RB Ser807/811 (D20B12) 1:1000, PDCD4 (D29C6) 1:1000, cleaved caspase 3 Asp175 (5A1E or rabbit polyclonal) 1:500, survivin (71G4B7) 1:1000 and p-S6K1 T389 (108D2) 1:1000, p-TSC2 S939 (rabbit polyclonal) 1:1000 obtained from Cell Signaling Technology.. Bcl-2 (N-19) 1:500, cyclin D1 (M-20) 1:500, p27Kip1 (C-19) 1:1000, cyclin A (C-19) 1:500, CDK4 (C-22) 1:1000, CDK6 (H-96) 1:500, and E2F1 (KH95) 1:500 were fom Santa Cruz. α-tubulin (CP06) 1:3000 was purchased from Calbiochem and GAPDH (rabbit polyclonal) 1:10000 from Trevigen.Bcl-2 (N-19) 1:500, cyclin D1 (M-20) 1:500, p27Kip1 (C-19) 1:1000, cyclin A (C-19) 1:500, CDK4 (C-22) 1:1000, CDK6 (H-96) 1:500, and E2F1 (KH95) 1:500 were fom Santa Cruz. α-tubulin (CP06) 1:3000 was purchased..

    Article Title: TSC1 promotes i NKT cell anergy and inhibits i NKT cell-mediated anti-tumor immunity
    Article Snippet: Fluorescence-conjugated anti-mouse TCRβ (H57-597), NK1.1 (PK136), CD44(IM7), B220(RA3-6B2), Gr-1(RB6-8C5), CD11c(N418), CD11b(M1/70), CD4 (GK1.5), CD8 (53-6.7), IFN-γ (XMG1.2), IL-4 (11B11), PD-1(RMP1-30), and ICOS(D10.G4.1) antibodies were purchased from BioLegend.Fluorescence-conjugated anti-mouse TCRβ (H57-597), NK1.1 (PK136), CD44(IM7), B220(RA3-6B2), Gr-1(RB6-8C5), CD11c(N418), CD11b(M1/70), CD4 (GK1.5), CD8 (53-6.7), IFN-γ (XMG1.2), IL-4 (11B11), PD-1(RMP1-30), and ICOS(D10.G4.1) antibodies were purchased from BioLegend.. Anti–phospho-p44/42 mitogen-activated protein kinase T202/Y204 (197G2), anti–phospho-S6 S240/244 (D68F8), anti–phospho-4E-BP1 T37/46 (236B4), anti–phospho-Akt S473 (193H12), TSC1 (D43E2), and TSC2 (D93F12) were purchased from Cell Signaling Technology, Inc. Anti–β-actin antibody was purchased from Sigma-Aldrich.. Cell surface staining was performed with 2% FBS-PBS.Cell surface staining was performed with 2% FBS-PBS.

    Article Title: TSC1 promotes i NKT cell anergy and inhibits i NKT cell-mediated anti-tumor immunity
    Article Snippet: CD11b(M1/70), CD4 (GK1.5), CD8 (53-6.7), IFN-γ (XMG1.2), IL-4 (11B11), PD-1(RMP1-30), and ICOS(D10.G4.1) antibodies were purchased from BioLegend. .. Anti–phospho-p44/42 mitogen-activated protein kinase T202/Y204 (197G2), anti–phospho-S6 S240/244 (D68F8), anti–phospho-4E-BP1 T37/46 (236B4), anti–phospho-Akt S473 (193H12), TSC1 (D43E2), and TSC2 (D93F12) were purchased from Cell Signaling Technology, Inc. Anti–β-actin antibody was purchased from Sigma-Aldrich. .. Cell surface staining was performed with 2% FBS-PBS.

    MANN-WHITNEY:

    Article Title: Trans‐gnetin H isolated from the seeds of Paeonia species induces autophagy via inhibiting mTORC1 signalling through AMPK activation
    Article Snippet: The WB protocol was the same as our previous work.The WB protocol was the same as our previous work.. The primary antibodies Actin (20536‐1‐AP), LC3II (18725‐1‐AP), p62 (18420‐1‐AP) were purchased from Proteintech; TFEB (37785 S), pT389‐S6K (9234 S), p‐S6 (4858 S), S6K (9202 S), S6 (2217 S), p‐AMPKα (50081 S), AMPKα (5832 S), Raptor (2280 S) and TSC2 (D93F12) were purchased from Cell Signaling Technology; Flag (db7002) and HA (db2603) were purchased from Hangzhou Bio Technolog.. The secondary antibodies were obtained from Sigma.The secondary antibodies were obtained from Sigma.

    Article Title: A translational program that suppresses metabolism to shield the genome
    Article Snippet: Blots were stripped and reprobed when looking at total versus modified proteins unless otherwise noted.. Antibodies (all 1:1000): GLUT1 (Novus Biologicals, # NB110-39113), PAI-1 (Proteintech, #13801-1-AP), Puromycin (3RH11) (Kerafast, #EQ0001), β-ACTIN (C4) (SantaCruz Biotechnology, #sc-47778), EIF5A (abcam, #ab32443), EIF3D (Proteintech, # 10219-1-AP), EIF3K (Proteintech, # 10640-1-AP), acetyl-EIF5A (Lys47) (Boster Bio, #P01727), CYR61 (Proteintech, #26689-1-AP), SDC4 (R&D Systems, #P31431), SIRT1 (Proteintech, #13161-1-AP), SIRT2 (Proteintech, #19655-1-AP), NFKB2 (Proteintech, #10409-2-AP), 4E-BP (Cell Signaling, #9452 S), Phospho-4E-BP (Ser65) (Cell Signaling, #9451 S), TSC2 (D93F12) (Cell Signaling, #4308S), gamma H2AX (Novus, NB100-384), H2AX (Abcam, Ab11175), CRM-1 (Novus, NB100-79802), XPO-4 (Abcam, ab133237), eIF5A2 (Thermo, #PA5-30770), Phospho-RAPTOR (Ser792) (Cell Signaling, #89146S), RAPTOR (Cell Signaling, #2280), Phospho-Tuberin/TSC2 (Ser1387) (Cell Signaling, #5584S), NDRG1 (Abcam, #ab37897), and Anti-hypusine (mAbHpu24) antibody was kindly provided by Genentech.. The eIF5A1 antibodies and siRNAs used in this study were specific to eIF5A1 and not eIF5A2 as described by the manufacturer and validated in Supplementary Fig. .

    Article Title: Identification of mechanically regulated phosphorylation sites on tuberin (TSC2) that control mechanistic target of rapamycin (mTOR) signaling
    Article Snippet: Images were quantified using ImageJ software (U.S. National Institutes of Health, Bethesda, MD; http://rsb.info.nih.gov/nih-image/).Images were quantified using ImageJ software (U.S. National Institutes of Health, Bethesda, MD; http://rsb.info.nih.gov/nih-image/).. Antibodies - P-p70(389) (1A5) (#9206, used to probe Myc-tagged P-p70), P-p70(389) (#9234, used to probe endogenous P-p70), total p70 (49D7) (#2708), Anti-P-Akt substrate (RxRxxS*/T*) (23C8D2) for detection of PTSC2 (#10001), P-JNK(Thr183/Tyr185) (98F2) (#4671), total JNK (#9252), total mTOR (#2972), total Rictor (D16H9) (#9476), P-PKB (Ser473) (#4060), P-PKB (Thr308) (#9275S), total PKB (#9272), P-S6 (Ser240/4) (#5364), P-S6 (Ser235/6) (#2211), total S6 (#2217S), total TSC1 (D43E2) (#6935), total TSC2 (D93F12) (#4308) were purchased from Cell Signaling Technology (Danvers, MA, USA).. Anti-puromycin (12D10) (#MABE343) was purchased from EMD Millipore (Billerica, MA, USA).Anti-puromycin (12D10) (#MABE343) was purchased from EMD Millipore (Billerica, MA, USA).

    Article Title: Chronic signaling via the metabolic checkpoint kinase mTORC1 induces macrophage granuloma formation and marks sarcoidosis progression
    Article Snippet: HRP-conjugated secondary antibodies (Bethyl) 1:10000 were applied for 1 hour at room temperature in 4% low-fat milk and proteins were visualized using ECL substrate (Thermo Scientific).HRP-conjugated secondary antibodies (Bethyl) 1:10000 were applied for 1 hour at room temperature in 4% low-fat milk and proteins were visualized using ECL substrate (Thermo Scientific).. Primary antibodies were TSC2 (D93F12) 1:1000, p-Akt S473 (D9E) 1:1000, 4E-BP1 (53H11) 1:1000, p-S6 S240/244 (rabbit polyclonal) 1:3000, S6 (54D2) 1:1000, p-Akt T308 (D25E6) 1:1000, pan-Akt (40D4) 1:1000, β-tubulin (9F3) 1:1000 , p-RB Ser807/811 (D20B12) 1:1000, PDCD4 (D29C6) 1:1000, cleaved caspase 3 Asp175 (5A1E or rabbit polyclonal) 1:500, survivin (71G4B7) 1:1000 and p-S6K1 T389 (108D2) 1:1000, p-TSC2 S939 (rabbit polyclonal) 1:1000 obtained from Cell Signaling Technology.. Bcl-2 (N-19) 1:500, cyclin D1 (M-20) 1:500, p27Kip1 (C-19) 1:1000, cyclin A (C-19) 1:500, CDK4 (C-22) 1:1000, CDK6 (H-96) 1:500, and E2F1 (KH95) 1:500 were fom Santa Cruz. α-tubulin (CP06) 1:3000 was purchased from Calbiochem and GAPDH (rabbit polyclonal) 1:10000 from Trevigen.Bcl-2 (N-19) 1:500, cyclin D1 (M-20) 1:500, p27Kip1 (C-19) 1:1000, cyclin A (C-19) 1:500, CDK4 (C-22) 1:1000, CDK6 (H-96) 1:500, and E2F1 (KH95) 1:500 were fom Santa Cruz. α-tubulin (CP06) 1:3000 was purchased..

    Article Title: TSC1 promotes i NKT cell anergy and inhibits i NKT cell-mediated anti-tumor immunity
    Article Snippet: Fluorescence-conjugated anti-mouse TCRβ (H57-597), NK1.1 (PK136), CD44(IM7), B220(RA3-6B2), Gr-1(RB6-8C5), CD11c(N418), CD11b(M1/70), CD4 (GK1.5), CD8 (53-6.7), IFN-γ (XMG1.2), IL-4 (11B11), PD-1(RMP1-30), and ICOS(D10.G4.1) antibodies were purchased from BioLegend.Fluorescence-conjugated anti-mouse TCRβ (H57-597), NK1.1 (PK136), CD44(IM7), B220(RA3-6B2), Gr-1(RB6-8C5), CD11c(N418), CD11b(M1/70), CD4 (GK1.5), CD8 (53-6.7), IFN-γ (XMG1.2), IL-4 (11B11), PD-1(RMP1-30), and ICOS(D10.G4.1) antibodies were purchased from BioLegend.. Anti–phospho-p44/42 mitogen-activated protein kinase T202/Y204 (197G2), anti–phospho-S6 S240/244 (D68F8), anti–phospho-4E-BP1 T37/46 (236B4), anti–phospho-Akt S473 (193H12), TSC1 (D43E2), and TSC2 (D93F12) were purchased from Cell Signaling Technology, Inc. Anti–β-actin antibody was purchased from Sigma-Aldrich.. Cell surface staining was performed with 2% FBS-PBS.Cell surface staining was performed with 2% FBS-PBS.

    Article Title: TSC1 promotes i NKT cell anergy and inhibits i NKT cell-mediated anti-tumor immunity
    Article Snippet: CD11b(M1/70), CD4 (GK1.5), CD8 (53-6.7), IFN-γ (XMG1.2), IL-4 (11B11), PD-1(RMP1-30), and ICOS(D10.G4.1) antibodies were purchased from BioLegend. .. Anti–phospho-p44/42 mitogen-activated protein kinase T202/Y204 (197G2), anti–phospho-S6 S240/244 (D68F8), anti–phospho-4E-BP1 T37/46 (236B4), anti–phospho-Akt S473 (193H12), TSC1 (D43E2), and TSC2 (D93F12) were purchased from Cell Signaling Technology, Inc. Anti–β-actin antibody was purchased from Sigma-Aldrich. .. Cell surface staining was performed with 2% FBS-PBS.

    Whisker Assay:

    Article Title: Trans‐gnetin H isolated from the seeds of Paeonia species induces autophagy via inhibiting mTORC1 signalling through AMPK activation
    Article Snippet: The WB protocol was the same as our previous work.The WB protocol was the same as our previous work.. The primary antibodies Actin (20536‐1‐AP), LC3II (18725‐1‐AP), p62 (18420‐1‐AP) were purchased from Proteintech; TFEB (37785 S), pT389‐S6K (9234 S), p‐S6 (4858 S), S6K (9202 S), S6 (2217 S), p‐AMPKα (50081 S), AMPKα (5832 S), Raptor (2280 S) and TSC2 (D93F12) were purchased from Cell Signaling Technology; Flag (db7002) and HA (db2603) were purchased from Hangzhou Bio Technolog.. The secondary antibodies were obtained from Sigma.The secondary antibodies were obtained from Sigma.

    Article Title: A translational program that suppresses metabolism to shield the genome
    Article Snippet: Blots were stripped and reprobed when looking at total versus modified proteins unless otherwise noted.. Antibodies (all 1:1000): GLUT1 (Novus Biologicals, # NB110-39113), PAI-1 (Proteintech, #13801-1-AP), Puromycin (3RH11) (Kerafast, #EQ0001), β-ACTIN (C4) (SantaCruz Biotechnology, #sc-47778), EIF5A (abcam, #ab32443), EIF3D (Proteintech, # 10219-1-AP), EIF3K (Proteintech, # 10640-1-AP), acetyl-EIF5A (Lys47) (Boster Bio, #P01727), CYR61 (Proteintech, #26689-1-AP), SDC4 (R&D Systems, #P31431), SIRT1 (Proteintech, #13161-1-AP), SIRT2 (Proteintech, #19655-1-AP), NFKB2 (Proteintech, #10409-2-AP), 4E-BP (Cell Signaling, #9452 S), Phospho-4E-BP (Ser65) (Cell Signaling, #9451 S), TSC2 (D93F12) (Cell Signaling, #4308S), gamma H2AX (Novus, NB100-384), H2AX (Abcam, Ab11175), CRM-1 (Novus, NB100-79802), XPO-4 (Abcam, ab133237), eIF5A2 (Thermo, #PA5-30770), Phospho-RAPTOR (Ser792) (Cell Signaling, #89146S), RAPTOR (Cell Signaling, #2280), Phospho-Tuberin/TSC2 (Ser1387) (Cell Signaling, #5584S), NDRG1 (Abcam, #ab37897), and Anti-hypusine (mAbHpu24) antibody was kindly provided by Genentech.. The eIF5A1 antibodies and siRNAs used in this study were specific to eIF5A1 and not eIF5A2 as described by the manufacturer and validated in Supplementary Fig. .

    Article Title: Identification of mechanically regulated phosphorylation sites on tuberin (TSC2) that control mechanistic target of rapamycin (mTOR) signaling
    Article Snippet: Images were quantified using ImageJ software (U.S. National Institutes of Health, Bethesda, MD; http://rsb.info.nih.gov/nih-image/).Images were quantified using ImageJ software (U.S. National Institutes of Health, Bethesda, MD; http://rsb.info.nih.gov/nih-image/).. Antibodies - P-p70(389) (1A5) (#9206, used to probe Myc-tagged P-p70), P-p70(389) (#9234, used to probe endogenous P-p70), total p70 (49D7) (#2708), Anti-P-Akt substrate (RxRxxS*/T*) (23C8D2) for detection of PTSC2 (#10001), P-JNK(Thr183/Tyr185) (98F2) (#4671), total JNK (#9252), total mTOR (#2972), total Rictor (D16H9) (#9476), P-PKB (Ser473) (#4060), P-PKB (Thr308) (#9275S), total PKB (#9272), P-S6 (Ser240/4) (#5364), P-S6 (Ser235/6) (#2211), total S6 (#2217S), total TSC1 (D43E2) (#6935), total TSC2 (D93F12) (#4308) were purchased from Cell Signaling Technology (Danvers, MA, USA).. Anti-puromycin (12D10) (#MABE343) was purchased from EMD Millipore (Billerica, MA, USA).Anti-puromycin (12D10) (#MABE343) was purchased from EMD Millipore (Billerica, MA, USA).

    Article Title: Chronic signaling via the metabolic checkpoint kinase mTORC1 induces macrophage granuloma formation and marks sarcoidosis progression
    Article Snippet: HRP-conjugated secondary antibodies (Bethyl) 1:10000 were applied for 1 hour at room temperature in 4% low-fat milk and proteins were visualized using ECL substrate (Thermo Scientific).HRP-conjugated secondary antibodies (Bethyl) 1:10000 were applied for 1 hour at room temperature in 4% low-fat milk and proteins were visualized using ECL substrate (Thermo Scientific).. Primary antibodies were TSC2 (D93F12) 1:1000, p-Akt S473 (D9E) 1:1000, 4E-BP1 (53H11) 1:1000, p-S6 S240/244 (rabbit polyclonal) 1:3000, S6 (54D2) 1:1000, p-Akt T308 (D25E6) 1:1000, pan-Akt (40D4) 1:1000, β-tubulin (9F3) 1:1000 , p-RB Ser807/811 (D20B12) 1:1000, PDCD4 (D29C6) 1:1000, cleaved caspase 3 Asp175 (5A1E or rabbit polyclonal) 1:500, survivin (71G4B7) 1:1000 and p-S6K1 T389 (108D2) 1:1000, p-TSC2 S939 (rabbit polyclonal) 1:1000 obtained from Cell Signaling Technology.. Bcl-2 (N-19) 1:500, cyclin D1 (M-20) 1:500, p27Kip1 (C-19) 1:1000, cyclin A (C-19) 1:500, CDK4 (C-22) 1:1000, CDK6 (H-96) 1:500, and E2F1 (KH95) 1:500 were fom Santa Cruz. α-tubulin (CP06) 1:3000 was purchased from Calbiochem and GAPDH (rabbit polyclonal) 1:10000 from Trevigen.Bcl-2 (N-19) 1:500, cyclin D1 (M-20) 1:500, p27Kip1 (C-19) 1:1000, cyclin A (C-19) 1:500, CDK4 (C-22) 1:1000, CDK6 (H-96) 1:500, and E2F1 (KH95) 1:500 were fom Santa Cruz. α-tubulin (CP06) 1:3000 was purchased..

    Article Title: TSC1 promotes i NKT cell anergy and inhibits i NKT cell-mediated anti-tumor immunity
    Article Snippet: Fluorescence-conjugated anti-mouse TCRβ (H57-597), NK1.1 (PK136), CD44(IM7), B220(RA3-6B2), Gr-1(RB6-8C5), CD11c(N418), CD11b(M1/70), CD4 (GK1.5), CD8 (53-6.7), IFN-γ (XMG1.2), IL-4 (11B11), PD-1(RMP1-30), and ICOS(D10.G4.1) antibodies were purchased from BioLegend.Fluorescence-conjugated anti-mouse TCRβ (H57-597), NK1.1 (PK136), CD44(IM7), B220(RA3-6B2), Gr-1(RB6-8C5), CD11c(N418), CD11b(M1/70), CD4 (GK1.5), CD8 (53-6.7), IFN-γ (XMG1.2), IL-4 (11B11), PD-1(RMP1-30), and ICOS(D10.G4.1) antibodies were purchased from BioLegend.. Anti–phospho-p44/42 mitogen-activated protein kinase T202/Y204 (197G2), anti–phospho-S6 S240/244 (D68F8), anti–phospho-4E-BP1 T37/46 (236B4), anti–phospho-Akt S473 (193H12), TSC1 (D43E2), and TSC2 (D93F12) were purchased from Cell Signaling Technology, Inc. Anti–β-actin antibody was purchased from Sigma-Aldrich.. Cell surface staining was performed with 2% FBS-PBS.Cell surface staining was performed with 2% FBS-PBS.

    Article Title: TSC1 promotes i NKT cell anergy and inhibits i NKT cell-mediated anti-tumor immunity
    Article Snippet: CD11b(M1/70), CD4 (GK1.5), CD8 (53-6.7), IFN-γ (XMG1.2), IL-4 (11B11), PD-1(RMP1-30), and ICOS(D10.G4.1) antibodies were purchased from BioLegend. .. Anti–phospho-p44/42 mitogen-activated protein kinase T202/Y204 (197G2), anti–phospho-S6 S240/244 (D68F8), anti–phospho-4E-BP1 T37/46 (236B4), anti–phospho-Akt S473 (193H12), TSC1 (D43E2), and TSC2 (D93F12) were purchased from Cell Signaling Technology, Inc. Anti–β-actin antibody was purchased from Sigma-Aldrich. .. Cell surface staining was performed with 2% FBS-PBS.

    Immunoprecipitation:

    Article Title: Trans‐gnetin H isolated from the seeds of Paeonia species induces autophagy via inhibiting mTORC1 signalling through AMPK activation
    Article Snippet: The WB protocol was the same as our previous work.The WB protocol was the same as our previous work.. The primary antibodies Actin (20536‐1‐AP), LC3II (18725‐1‐AP), p62 (18420‐1‐AP) were purchased from Proteintech; TFEB (37785 S), pT389‐S6K (9234 S), p‐S6 (4858 S), S6K (9202 S), S6 (2217 S), p‐AMPKα (50081 S), AMPKα (5832 S), Raptor (2280 S) and TSC2 (D93F12) were purchased from Cell Signaling Technology; Flag (db7002) and HA (db2603) were purchased from Hangzhou Bio Technolog.. The secondary antibodies were obtained from Sigma.The secondary antibodies were obtained from Sigma.

    Article Title: A translational program that suppresses metabolism to shield the genome
    Article Snippet: Blots were stripped and reprobed when looking at total versus modified proteins unless otherwise noted.. Antibodies (all 1:1000): GLUT1 (Novus Biologicals, # NB110-39113), PAI-1 (Proteintech, #13801-1-AP), Puromycin (3RH11) (Kerafast, #EQ0001), β-ACTIN (C4) (SantaCruz Biotechnology, #sc-47778), EIF5A (abcam, #ab32443), EIF3D (Proteintech, # 10219-1-AP), EIF3K (Proteintech, # 10640-1-AP), acetyl-EIF5A (Lys47) (Boster Bio, #P01727), CYR61 (Proteintech, #26689-1-AP), SDC4 (R&D Systems, #P31431), SIRT1 (Proteintech, #13161-1-AP), SIRT2 (Proteintech, #19655-1-AP), NFKB2 (Proteintech, #10409-2-AP), 4E-BP (Cell Signaling, #9452 S), Phospho-4E-BP (Ser65) (Cell Signaling, #9451 S), TSC2 (D93F12) (Cell Signaling, #4308S), gamma H2AX (Novus, NB100-384), H2AX (Abcam, Ab11175), CRM-1 (Novus, NB100-79802), XPO-4 (Abcam, ab133237), eIF5A2 (Thermo, #PA5-30770), Phospho-RAPTOR (Ser792) (Cell Signaling, #89146S), RAPTOR (Cell Signaling, #2280), Phospho-Tuberin/TSC2 (Ser1387) (Cell Signaling, #5584S), NDRG1 (Abcam, #ab37897), and Anti-hypusine (mAbHpu24) antibody was kindly provided by Genentech.. The eIF5A1 antibodies and siRNAs used in this study were specific to eIF5A1 and not eIF5A2 as described by the manufacturer and validated in Supplementary Fig. .

    Article Title: Identification of mechanically regulated phosphorylation sites on tuberin (TSC2) that control mechanistic target of rapamycin (mTOR) signaling
    Article Snippet: Images were quantified using ImageJ software (U.S. National Institutes of Health, Bethesda, MD; http://rsb.info.nih.gov/nih-image/).Images were quantified using ImageJ software (U.S. National Institutes of Health, Bethesda, MD; http://rsb.info.nih.gov/nih-image/).. Antibodies - P-p70(389) (1A5) (#9206, used to probe Myc-tagged P-p70), P-p70(389) (#9234, used to probe endogenous P-p70), total p70 (49D7) (#2708), Anti-P-Akt substrate (RxRxxS*/T*) (23C8D2) for detection of PTSC2 (#10001), P-JNK(Thr183/Tyr185) (98F2) (#4671), total JNK (#9252), total mTOR (#2972), total Rictor (D16H9) (#9476), P-PKB (Ser473) (#4060), P-PKB (Thr308) (#9275S), total PKB (#9272), P-S6 (Ser240/4) (#5364), P-S6 (Ser235/6) (#2211), total S6 (#2217S), total TSC1 (D43E2) (#6935), total TSC2 (D93F12) (#4308) were purchased from Cell Signaling Technology (Danvers, MA, USA).. Anti-puromycin (12D10) (#MABE343) was purchased from EMD Millipore (Billerica, MA, USA).Anti-puromycin (12D10) (#MABE343) was purchased from EMD Millipore (Billerica, MA, USA).

    Article Title: Chronic signaling via the metabolic checkpoint kinase mTORC1 induces macrophage granuloma formation and marks sarcoidosis progression
    Article Snippet: HRP-conjugated secondary antibodies (Bethyl) 1:10000 were applied for 1 hour at room temperature in 4% low-fat milk and proteins were visualized using ECL substrate (Thermo Scientific).HRP-conjugated secondary antibodies (Bethyl) 1:10000 were applied for 1 hour at room temperature in 4% low-fat milk and proteins were visualized using ECL substrate (Thermo Scientific).. Primary antibodies were TSC2 (D93F12) 1:1000, p-Akt S473 (D9E) 1:1000, 4E-BP1 (53H11) 1:1000, p-S6 S240/244 (rabbit polyclonal) 1:3000, S6 (54D2) 1:1000, p-Akt T308 (D25E6) 1:1000, pan-Akt (40D4) 1:1000, β-tubulin (9F3) 1:1000 , p-RB Ser807/811 (D20B12) 1:1000, PDCD4 (D29C6) 1:1000, cleaved caspase 3 Asp175 (5A1E or rabbit polyclonal) 1:500, survivin (71G4B7) 1:1000 and p-S6K1 T389 (108D2) 1:1000, p-TSC2 S939 (rabbit polyclonal) 1:1000 obtained from Cell Signaling Technology.. Bcl-2 (N-19) 1:500, cyclin D1 (M-20) 1:500, p27Kip1 (C-19) 1:1000, cyclin A (C-19) 1:500, CDK4 (C-22) 1:1000, CDK6 (H-96) 1:500, and E2F1 (KH95) 1:500 were fom Santa Cruz. α-tubulin (CP06) 1:3000 was purchased from Calbiochem and GAPDH (rabbit polyclonal) 1:10000 from Trevigen.Bcl-2 (N-19) 1:500, cyclin D1 (M-20) 1:500, p27Kip1 (C-19) 1:1000, cyclin A (C-19) 1:500, CDK4 (C-22) 1:1000, CDK6 (H-96) 1:500, and E2F1 (KH95) 1:500 were fom Santa Cruz. α-tubulin (CP06) 1:3000 was purchased..

    Article Title: TSC1 promotes i NKT cell anergy and inhibits i NKT cell-mediated anti-tumor immunity
    Article Snippet: Fluorescence-conjugated anti-mouse TCRβ (H57-597), NK1.1 (PK136), CD44(IM7), B220(RA3-6B2), Gr-1(RB6-8C5), CD11c(N418), CD11b(M1/70), CD4 (GK1.5), CD8 (53-6.7), IFN-γ (XMG1.2), IL-4 (11B11), PD-1(RMP1-30), and ICOS(D10.G4.1) antibodies were purchased from BioLegend.Fluorescence-conjugated anti-mouse TCRβ (H57-597), NK1.1 (PK136), CD44(IM7), B220(RA3-6B2), Gr-1(RB6-8C5), CD11c(N418), CD11b(M1/70), CD4 (GK1.5), CD8 (53-6.7), IFN-γ (XMG1.2), IL-4 (11B11), PD-1(RMP1-30), and ICOS(D10.G4.1) antibodies were purchased from BioLegend.. Anti–phospho-p44/42 mitogen-activated protein kinase T202/Y204 (197G2), anti–phospho-S6 S240/244 (D68F8), anti–phospho-4E-BP1 T37/46 (236B4), anti–phospho-Akt S473 (193H12), TSC1 (D43E2), and TSC2 (D93F12) were purchased from Cell Signaling Technology, Inc. Anti–β-actin antibody was purchased from Sigma-Aldrich.. Cell surface staining was performed with 2% FBS-PBS.Cell surface staining was performed with 2% FBS-PBS.

    Article Title: TSC1 promotes i NKT cell anergy and inhibits i NKT cell-mediated anti-tumor immunity
    Article Snippet: CD11b(M1/70), CD4 (GK1.5), CD8 (53-6.7), IFN-γ (XMG1.2), IL-4 (11B11), PD-1(RMP1-30), and ICOS(D10.G4.1) antibodies were purchased from BioLegend. .. Anti–phospho-p44/42 mitogen-activated protein kinase T202/Y204 (197G2), anti–phospho-S6 S240/244 (D68F8), anti–phospho-4E-BP1 T37/46 (236B4), anti–phospho-Akt S473 (193H12), TSC1 (D43E2), and TSC2 (D93F12) were purchased from Cell Signaling Technology, Inc. Anti–β-actin antibody was purchased from Sigma-Aldrich. .. Cell surface staining was performed with 2% FBS-PBS.

    Control:

    Article Title: Trans‐gnetin H isolated from the seeds of Paeonia species induces autophagy via inhibiting mTORC1 signalling through AMPK activation
    Article Snippet: The WB protocol was the same as our previous work.The WB protocol was the same as our previous work.. The primary antibodies Actin (20536‐1‐AP), LC3II (18725‐1‐AP), p62 (18420‐1‐AP) were purchased from Proteintech; TFEB (37785 S), pT389‐S6K (9234 S), p‐S6 (4858 S), S6K (9202 S), S6 (2217 S), p‐AMPKα (50081 S), AMPKα (5832 S), Raptor (2280 S) and TSC2 (D93F12) were purchased from Cell Signaling Technology; Flag (db7002) and HA (db2603) were purchased from Hangzhou Bio Technolog.. The secondary antibodies were obtained from Sigma.The secondary antibodies were obtained from Sigma.

    Article Title: A translational program that suppresses metabolism to shield the genome
    Article Snippet: Blots were stripped and reprobed when looking at total versus modified proteins unless otherwise noted.. Antibodies (all 1:1000): GLUT1 (Novus Biologicals, # NB110-39113), PAI-1 (Proteintech, #13801-1-AP), Puromycin (3RH11) (Kerafast, #EQ0001), β-ACTIN (C4) (SantaCruz Biotechnology, #sc-47778), EIF5A (abcam, #ab32443), EIF3D (Proteintech, # 10219-1-AP), EIF3K (Proteintech, # 10640-1-AP), acetyl-EIF5A (Lys47) (Boster Bio, #P01727), CYR61 (Proteintech, #26689-1-AP), SDC4 (R&D Systems, #P31431), SIRT1 (Proteintech, #13161-1-AP), SIRT2 (Proteintech, #19655-1-AP), NFKB2 (Proteintech, #10409-2-AP), 4E-BP (Cell Signaling, #9452 S), Phospho-4E-BP (Ser65) (Cell Signaling, #9451 S), TSC2 (D93F12) (Cell Signaling, #4308S), gamma H2AX (Novus, NB100-384), H2AX (Abcam, Ab11175), CRM-1 (Novus, NB100-79802), XPO-4 (Abcam, ab133237), eIF5A2 (Thermo, #PA5-30770), Phospho-RAPTOR (Ser792) (Cell Signaling, #89146S), RAPTOR (Cell Signaling, #2280), Phospho-Tuberin/TSC2 (Ser1387) (Cell Signaling, #5584S), NDRG1 (Abcam, #ab37897), and Anti-hypusine (mAbHpu24) antibody was kindly provided by Genentech.. The eIF5A1 antibodies and siRNAs used in this study were specific to eIF5A1 and not eIF5A2 as described by the manufacturer and validated in Supplementary Fig. .

    Article Title: Identification of mechanically regulated phosphorylation sites on tuberin (TSC2) that control mechanistic target of rapamycin (mTOR) signaling
    Article Snippet: Images were quantified using ImageJ software (U.S. National Institutes of Health, Bethesda, MD; http://rsb.info.nih.gov/nih-image/).Images were quantified using ImageJ software (U.S. National Institutes of Health, Bethesda, MD; http://rsb.info.nih.gov/nih-image/).. Antibodies - P-p70(389) (1A5) (#9206, used to probe Myc-tagged P-p70), P-p70(389) (#9234, used to probe endogenous P-p70), total p70 (49D7) (#2708), Anti-P-Akt substrate (RxRxxS*/T*) (23C8D2) for detection of PTSC2 (#10001), P-JNK(Thr183/Tyr185) (98F2) (#4671), total JNK (#9252), total mTOR (#2972), total Rictor (D16H9) (#9476), P-PKB (Ser473) (#4060), P-PKB (Thr308) (#9275S), total PKB (#9272), P-S6 (Ser240/4) (#5364), P-S6 (Ser235/6) (#2211), total S6 (#2217S), total TSC1 (D43E2) (#6935), total TSC2 (D93F12) (#4308) were purchased from Cell Signaling Technology (Danvers, MA, USA).. Anti-puromycin (12D10) (#MABE343) was purchased from EMD Millipore (Billerica, MA, USA).Anti-puromycin (12D10) (#MABE343) was purchased from EMD Millipore (Billerica, MA, USA).

    Article Title: Chronic signaling via the metabolic checkpoint kinase mTORC1 induces macrophage granuloma formation and marks sarcoidosis progression
    Article Snippet: HRP-conjugated secondary antibodies (Bethyl) 1:10000 were applied for 1 hour at room temperature in 4% low-fat milk and proteins were visualized using ECL substrate (Thermo Scientific).HRP-conjugated secondary antibodies (Bethyl) 1:10000 were applied for 1 hour at room temperature in 4% low-fat milk and proteins were visualized using ECL substrate (Thermo Scientific).. Primary antibodies were TSC2 (D93F12) 1:1000, p-Akt S473 (D9E) 1:1000, 4E-BP1 (53H11) 1:1000, p-S6 S240/244 (rabbit polyclonal) 1:3000, S6 (54D2) 1:1000, p-Akt T308 (D25E6) 1:1000, pan-Akt (40D4) 1:1000, β-tubulin (9F3) 1:1000 , p-RB Ser807/811 (D20B12) 1:1000, PDCD4 (D29C6) 1:1000, cleaved caspase 3 Asp175 (5A1E or rabbit polyclonal) 1:500, survivin (71G4B7) 1:1000 and p-S6K1 T389 (108D2) 1:1000, p-TSC2 S939 (rabbit polyclonal) 1:1000 obtained from Cell Signaling Technology.. Bcl-2 (N-19) 1:500, cyclin D1 (M-20) 1:500, p27Kip1 (C-19) 1:1000, cyclin A (C-19) 1:500, CDK4 (C-22) 1:1000, CDK6 (H-96) 1:500, and E2F1 (KH95) 1:500 were fom Santa Cruz. α-tubulin (CP06) 1:3000 was purchased from Calbiochem and GAPDH (rabbit polyclonal) 1:10000 from Trevigen.Bcl-2 (N-19) 1:500, cyclin D1 (M-20) 1:500, p27Kip1 (C-19) 1:1000, cyclin A (C-19) 1:500, CDK4 (C-22) 1:1000, CDK6 (H-96) 1:500, and E2F1 (KH95) 1:500 were fom Santa Cruz. α-tubulin (CP06) 1:3000 was purchased..

    Article Title: TSC1 promotes i NKT cell anergy and inhibits i NKT cell-mediated anti-tumor immunity
    Article Snippet: Fluorescence-conjugated anti-mouse TCRβ (H57-597), NK1.1 (PK136), CD44(IM7), B220(RA3-6B2), Gr-1(RB6-8C5), CD11c(N418), CD11b(M1/70), CD4 (GK1.5), CD8 (53-6.7), IFN-γ (XMG1.2), IL-4 (11B11), PD-1(RMP1-30), and ICOS(D10.G4.1) antibodies were purchased from BioLegend.Fluorescence-conjugated anti-mouse TCRβ (H57-597), NK1.1 (PK136), CD44(IM7), B220(RA3-6B2), Gr-1(RB6-8C5), CD11c(N418), CD11b(M1/70), CD4 (GK1.5), CD8 (53-6.7), IFN-γ (XMG1.2), IL-4 (11B11), PD-1(RMP1-30), and ICOS(D10.G4.1) antibodies were purchased from BioLegend.. Anti–phospho-p44/42 mitogen-activated protein kinase T202/Y204 (197G2), anti–phospho-S6 S240/244 (D68F8), anti–phospho-4E-BP1 T37/46 (236B4), anti–phospho-Akt S473 (193H12), TSC1 (D43E2), and TSC2 (D93F12) were purchased from Cell Signaling Technology, Inc. Anti–β-actin antibody was purchased from Sigma-Aldrich.. Cell surface staining was performed with 2% FBS-PBS.Cell surface staining was performed with 2% FBS-PBS.

    Article Title: TSC1 promotes i NKT cell anergy and inhibits i NKT cell-mediated anti-tumor immunity
    Article Snippet: CD11b(M1/70), CD4 (GK1.5), CD8 (53-6.7), IFN-γ (XMG1.2), IL-4 (11B11), PD-1(RMP1-30), and ICOS(D10.G4.1) antibodies were purchased from BioLegend. .. Anti–phospho-p44/42 mitogen-activated protein kinase T202/Y204 (197G2), anti–phospho-S6 S240/244 (D68F8), anti–phospho-4E-BP1 T37/46 (236B4), anti–phospho-Akt S473 (193H12), TSC1 (D43E2), and TSC2 (D93F12) were purchased from Cell Signaling Technology, Inc. Anti–β-actin antibody was purchased from Sigma-Aldrich. .. Cell surface staining was performed with 2% FBS-PBS.

    In Situ Hybridization:

    Article Title: Trans‐gnetin H isolated from the seeds of Paeonia species induces autophagy via inhibiting mTORC1 signalling through AMPK activation
    Article Snippet: The WB protocol was the same as our previous work.The WB protocol was the same as our previous work.. The primary antibodies Actin (20536‐1‐AP), LC3II (18725‐1‐AP), p62 (18420‐1‐AP) were purchased from Proteintech; TFEB (37785 S), pT389‐S6K (9234 S), p‐S6 (4858 S), S6K (9202 S), S6 (2217 S), p‐AMPKα (50081 S), AMPKα (5832 S), Raptor (2280 S) and TSC2 (D93F12) were purchased from Cell Signaling Technology; Flag (db7002) and HA (db2603) were purchased from Hangzhou Bio Technolog.. The secondary antibodies were obtained from Sigma.The secondary antibodies were obtained from Sigma.

    Article Title: A translational program that suppresses metabolism to shield the genome
    Article Snippet: Blots were stripped and reprobed when looking at total versus modified proteins unless otherwise noted.. Antibodies (all 1:1000): GLUT1 (Novus Biologicals, # NB110-39113), PAI-1 (Proteintech, #13801-1-AP), Puromycin (3RH11) (Kerafast, #EQ0001), β-ACTIN (C4) (SantaCruz Biotechnology, #sc-47778), EIF5A (abcam, #ab32443), EIF3D (Proteintech, # 10219-1-AP), EIF3K (Proteintech, # 10640-1-AP), acetyl-EIF5A (Lys47) (Boster Bio, #P01727), CYR61 (Proteintech, #26689-1-AP), SDC4 (R&D Systems, #P31431), SIRT1 (Proteintech, #13161-1-AP), SIRT2 (Proteintech, #19655-1-AP), NFKB2 (Proteintech, #10409-2-AP), 4E-BP (Cell Signaling, #9452 S), Phospho-4E-BP (Ser65) (Cell Signaling, #9451 S), TSC2 (D93F12) (Cell Signaling, #4308S), gamma H2AX (Novus, NB100-384), H2AX (Abcam, Ab11175), CRM-1 (Novus, NB100-79802), XPO-4 (Abcam, ab133237), eIF5A2 (Thermo, #PA5-30770), Phospho-RAPTOR (Ser792) (Cell Signaling, #89146S), RAPTOR (Cell Signaling, #2280), Phospho-Tuberin/TSC2 (Ser1387) (Cell Signaling, #5584S), NDRG1 (Abcam, #ab37897), and Anti-hypusine (mAbHpu24) antibody was kindly provided by Genentech.. The eIF5A1 antibodies and siRNAs used in this study were specific to eIF5A1 and not eIF5A2 as described by the manufacturer and validated in Supplementary Fig. .

    Article Title: Identification of mechanically regulated phosphorylation sites on tuberin (TSC2) that control mechanistic target of rapamycin (mTOR) signaling
    Article Snippet: Images were quantified using ImageJ software (U.S. National Institutes of Health, Bethesda, MD; http://rsb.info.nih.gov/nih-image/).Images were quantified using ImageJ software (U.S. National Institutes of Health, Bethesda, MD; http://rsb.info.nih.gov/nih-image/).. Antibodies - P-p70(389) (1A5) (#9206, used to probe Myc-tagged P-p70), P-p70(389) (#9234, used to probe endogenous P-p70), total p70 (49D7) (#2708), Anti-P-Akt substrate (RxRxxS*/T*) (23C8D2) for detection of PTSC2 (#10001), P-JNK(Thr183/Tyr185) (98F2) (#4671), total JNK (#9252), total mTOR (#2972), total Rictor (D16H9) (#9476), P-PKB (Ser473) (#4060), P-PKB (Thr308) (#9275S), total PKB (#9272), P-S6 (Ser240/4) (#5364), P-S6 (Ser235/6) (#2211), total S6 (#2217S), total TSC1 (D43E2) (#6935), total TSC2 (D93F12) (#4308) were purchased from Cell Signaling Technology (Danvers, MA, USA).. Anti-puromycin (12D10) (#MABE343) was purchased from EMD Millipore (Billerica, MA, USA).Anti-puromycin (12D10) (#MABE343) was purchased from EMD Millipore (Billerica, MA, USA).

    Article Title: Chronic signaling via the metabolic checkpoint kinase mTORC1 induces macrophage granuloma formation and marks sarcoidosis progression
    Article Snippet: HRP-conjugated secondary antibodies (Bethyl) 1:10000 were applied for 1 hour at room temperature in 4% low-fat milk and proteins were visualized using ECL substrate (Thermo Scientific).HRP-conjugated secondary antibodies (Bethyl) 1:10000 were applied for 1 hour at room temperature in 4% low-fat milk and proteins were visualized using ECL substrate (Thermo Scientific).. Primary antibodies were TSC2 (D93F12) 1:1000, p-Akt S473 (D9E) 1:1000, 4E-BP1 (53H11) 1:1000, p-S6 S240/244 (rabbit polyclonal) 1:3000, S6 (54D2) 1:1000, p-Akt T308 (D25E6) 1:1000, pan-Akt (40D4) 1:1000, β-tubulin (9F3) 1:1000 , p-RB Ser807/811 (D20B12) 1:1000, PDCD4 (D29C6) 1:1000, cleaved caspase 3 Asp175 (5A1E or rabbit polyclonal) 1:500, survivin (71G4B7) 1:1000 and p-S6K1 T389 (108D2) 1:1000, p-TSC2 S939 (rabbit polyclonal) 1:1000 obtained from Cell Signaling Technology.. Bcl-2 (N-19) 1:500, cyclin D1 (M-20) 1:500, p27Kip1 (C-19) 1:1000, cyclin A (C-19) 1:500, CDK4 (C-22) 1:1000, CDK6 (H-96) 1:500, and E2F1 (KH95) 1:500 were fom Santa Cruz. α-tubulin (CP06) 1:3000 was purchased from Calbiochem and GAPDH (rabbit polyclonal) 1:10000 from Trevigen.Bcl-2 (N-19) 1:500, cyclin D1 (M-20) 1:500, p27Kip1 (C-19) 1:1000, cyclin A (C-19) 1:500, CDK4 (C-22) 1:1000, CDK6 (H-96) 1:500, and E2F1 (KH95) 1:500 were fom Santa Cruz. α-tubulin (CP06) 1:3000 was purchased..

    Article Title: TSC1 promotes i NKT cell anergy and inhibits i NKT cell-mediated anti-tumor immunity
    Article Snippet: Fluorescence-conjugated anti-mouse TCRβ (H57-597), NK1.1 (PK136), CD44(IM7), B220(RA3-6B2), Gr-1(RB6-8C5), CD11c(N418), CD11b(M1/70), CD4 (GK1.5), CD8 (53-6.7), IFN-γ (XMG1.2), IL-4 (11B11), PD-1(RMP1-30), and ICOS(D10.G4.1) antibodies were purchased from BioLegend.Fluorescence-conjugated anti-mouse TCRβ (H57-597), NK1.1 (PK136), CD44(IM7), B220(RA3-6B2), Gr-1(RB6-8C5), CD11c(N418), CD11b(M1/70), CD4 (GK1.5), CD8 (53-6.7), IFN-γ (XMG1.2), IL-4 (11B11), PD-1(RMP1-30), and ICOS(D10.G4.1) antibodies were purchased from BioLegend.. Anti–phospho-p44/42 mitogen-activated protein kinase T202/Y204 (197G2), anti–phospho-S6 S240/244 (D68F8), anti–phospho-4E-BP1 T37/46 (236B4), anti–phospho-Akt S473 (193H12), TSC1 (D43E2), and TSC2 (D93F12) were purchased from Cell Signaling Technology, Inc. Anti–β-actin antibody was purchased from Sigma-Aldrich.. Cell surface staining was performed with 2% FBS-PBS.Cell surface staining was performed with 2% FBS-PBS.

    Article Title: TSC1 promotes i NKT cell anergy and inhibits i NKT cell-mediated anti-tumor immunity
    Article Snippet: CD11b(M1/70), CD4 (GK1.5), CD8 (53-6.7), IFN-γ (XMG1.2), IL-4 (11B11), PD-1(RMP1-30), and ICOS(D10.G4.1) antibodies were purchased from BioLegend. .. Anti–phospho-p44/42 mitogen-activated protein kinase T202/Y204 (197G2), anti–phospho-S6 S240/244 (D68F8), anti–phospho-4E-BP1 T37/46 (236B4), anti–phospho-Akt S473 (193H12), TSC1 (D43E2), and TSC2 (D93F12) were purchased from Cell Signaling Technology, Inc. Anti–β-actin antibody was purchased from Sigma-Aldrich. .. Cell surface staining was performed with 2% FBS-PBS.

    Knock-Out:

    Article Title: Trans‐gnetin H isolated from the seeds of Paeonia species induces autophagy via inhibiting mTORC1 signalling through AMPK activation
    Article Snippet: The WB protocol was the same as our previous work.The WB protocol was the same as our previous work.. The primary antibodies Actin (20536‐1‐AP), LC3II (18725‐1‐AP), p62 (18420‐1‐AP) were purchased from Proteintech; TFEB (37785 S), pT389‐S6K (9234 S), p‐S6 (4858 S), S6K (9202 S), S6 (2217 S), p‐AMPKα (50081 S), AMPKα (5832 S), Raptor (2280 S) and TSC2 (D93F12) were purchased from Cell Signaling Technology; Flag (db7002) and HA (db2603) were purchased from Hangzhou Bio Technolog.. The secondary antibodies were obtained from Sigma.The secondary antibodies were obtained from Sigma.

    Article Title: A translational program that suppresses metabolism to shield the genome
    Article Snippet: Blots were stripped and reprobed when looking at total versus modified proteins unless otherwise noted.. Antibodies (all 1:1000): GLUT1 (Novus Biologicals, # NB110-39113), PAI-1 (Proteintech, #13801-1-AP), Puromycin (3RH11) (Kerafast, #EQ0001), β-ACTIN (C4) (SantaCruz Biotechnology, #sc-47778), EIF5A (abcam, #ab32443), EIF3D (Proteintech, # 10219-1-AP), EIF3K (Proteintech, # 10640-1-AP), acetyl-EIF5A (Lys47) (Boster Bio, #P01727), CYR61 (Proteintech, #26689-1-AP), SDC4 (R&D Systems, #P31431), SIRT1 (Proteintech, #13161-1-AP), SIRT2 (Proteintech, #19655-1-AP), NFKB2 (Proteintech, #10409-2-AP), 4E-BP (Cell Signaling, #9452 S), Phospho-4E-BP (Ser65) (Cell Signaling, #9451 S), TSC2 (D93F12) (Cell Signaling, #4308S), gamma H2AX (Novus, NB100-384), H2AX (Abcam, Ab11175), CRM-1 (Novus, NB100-79802), XPO-4 (Abcam, ab133237), eIF5A2 (Thermo, #PA5-30770), Phospho-RAPTOR (Ser792) (Cell Signaling, #89146S), RAPTOR (Cell Signaling, #2280), Phospho-Tuberin/TSC2 (Ser1387) (Cell Signaling, #5584S), NDRG1 (Abcam, #ab37897), and Anti-hypusine (mAbHpu24) antibody was kindly provided by Genentech.. The eIF5A1 antibodies and siRNAs used in this study were specific to eIF5A1 and not eIF5A2 as described by the manufacturer and validated in Supplementary Fig. .

    Article Title: Identification of mechanically regulated phosphorylation sites on tuberin (TSC2) that control mechanistic target of rapamycin (mTOR) signaling
    Article Snippet: Images were quantified using ImageJ software (U.S. National Institutes of Health, Bethesda, MD; http://rsb.info.nih.gov/nih-image/).Images were quantified using ImageJ software (U.S. National Institutes of Health, Bethesda, MD; http://rsb.info.nih.gov/nih-image/).. Antibodies - P-p70(389) (1A5) (#9206, used to probe Myc-tagged P-p70), P-p70(389) (#9234, used to probe endogenous P-p70), total p70 (49D7) (#2708), Anti-P-Akt substrate (RxRxxS*/T*) (23C8D2) for detection of PTSC2 (#10001), P-JNK(Thr183/Tyr185) (98F2) (#4671), total JNK (#9252), total mTOR (#2972), total Rictor (D16H9) (#9476), P-PKB (Ser473) (#4060), P-PKB (Thr308) (#9275S), total PKB (#9272), P-S6 (Ser240/4) (#5364), P-S6 (Ser235/6) (#2211), total S6 (#2217S), total TSC1 (D43E2) (#6935), total TSC2 (D93F12) (#4308) were purchased from Cell Signaling Technology (Danvers, MA, USA).. Anti-puromycin (12D10) (#MABE343) was purchased from EMD Millipore (Billerica, MA, USA).Anti-puromycin (12D10) (#MABE343) was purchased from EMD Millipore (Billerica, MA, USA).

    Article Title: Chronic signaling via the metabolic checkpoint kinase mTORC1 induces macrophage granuloma formation and marks sarcoidosis progression
    Article Snippet: HRP-conjugated secondary antibodies (Bethyl) 1:10000 were applied for 1 hour at room temperature in 4% low-fat milk and proteins were visualized using ECL substrate (Thermo Scientific).HRP-conjugated secondary antibodies (Bethyl) 1:10000 were applied for 1 hour at room temperature in 4% low-fat milk and proteins were visualized using ECL substrate (Thermo Scientific).. Primary antibodies were TSC2 (D93F12) 1:1000, p-Akt S473 (D9E) 1:1000, 4E-BP1 (53H11) 1:1000, p-S6 S240/244 (rabbit polyclonal) 1:3000, S6 (54D2) 1:1000, p-Akt T308 (D25E6) 1:1000, pan-Akt (40D4) 1:1000, β-tubulin (9F3) 1:1000 , p-RB Ser807/811 (D20B12) 1:1000, PDCD4 (D29C6) 1:1000, cleaved caspase 3 Asp175 (5A1E or rabbit polyclonal) 1:500, survivin (71G4B7) 1:1000 and p-S6K1 T389 (108D2) 1:1000, p-TSC2 S939 (rabbit polyclonal) 1:1000 obtained from Cell Signaling Technology.. Bcl-2 (N-19) 1:500, cyclin D1 (M-20) 1:500, p27Kip1 (C-19) 1:1000, cyclin A (C-19) 1:500, CDK4 (C-22) 1:1000, CDK6 (H-96) 1:500, and E2F1 (KH95) 1:500 were fom Santa Cruz. α-tubulin (CP06) 1:3000 was purchased from Calbiochem and GAPDH (rabbit polyclonal) 1:10000 from Trevigen.Bcl-2 (N-19) 1:500, cyclin D1 (M-20) 1:500, p27Kip1 (C-19) 1:1000, cyclin A (C-19) 1:500, CDK4 (C-22) 1:1000, CDK6 (H-96) 1:500, and E2F1 (KH95) 1:500 were fom Santa Cruz. α-tubulin (CP06) 1:3000 was purchased..

    Article Title: TSC1 promotes i NKT cell anergy and inhibits i NKT cell-mediated anti-tumor immunity
    Article Snippet: Fluorescence-conjugated anti-mouse TCRβ (H57-597), NK1.1 (PK136), CD44(IM7), B220(RA3-6B2), Gr-1(RB6-8C5), CD11c(N418), CD11b(M1/70), CD4 (GK1.5), CD8 (53-6.7), IFN-γ (XMG1.2), IL-4 (11B11), PD-1(RMP1-30), and ICOS(D10.G4.1) antibodies were purchased from BioLegend.Fluorescence-conjugated anti-mouse TCRβ (H57-597), NK1.1 (PK136), CD44(IM7), B220(RA3-6B2), Gr-1(RB6-8C5), CD11c(N418), CD11b(M1/70), CD4 (GK1.5), CD8 (53-6.7), IFN-γ (XMG1.2), IL-4 (11B11), PD-1(RMP1-30), and ICOS(D10.G4.1) antibodies were purchased from BioLegend.. Anti–phospho-p44/42 mitogen-activated protein kinase T202/Y204 (197G2), anti–phospho-S6 S240/244 (D68F8), anti–phospho-4E-BP1 T37/46 (236B4), anti–phospho-Akt S473 (193H12), TSC1 (D43E2), and TSC2 (D93F12) were purchased from Cell Signaling Technology, Inc. Anti–β-actin antibody was purchased from Sigma-Aldrich.. Cell surface staining was performed with 2% FBS-PBS.Cell surface staining was performed with 2% FBS-PBS.

    Article Title: TSC1 promotes i NKT cell anergy and inhibits i NKT cell-mediated anti-tumor immunity
    Article Snippet: CD11b(M1/70), CD4 (GK1.5), CD8 (53-6.7), IFN-γ (XMG1.2), IL-4 (11B11), PD-1(RMP1-30), and ICOS(D10.G4.1) antibodies were purchased from BioLegend. .. Anti–phospho-p44/42 mitogen-activated protein kinase T202/Y204 (197G2), anti–phospho-S6 S240/244 (D68F8), anti–phospho-4E-BP1 T37/46 (236B4), anti–phospho-Akt S473 (193H12), TSC1 (D43E2), and TSC2 (D93F12) were purchased from Cell Signaling Technology, Inc. Anti–β-actin antibody was purchased from Sigma-Aldrich. .. Cell surface staining was performed with 2% FBS-PBS.

    Muscles:

    Article Title: Trans‐gnetin H isolated from the seeds of Paeonia species induces autophagy via inhibiting mTORC1 signalling through AMPK activation
    Article Snippet: The WB protocol was the same as our previous work.The WB protocol was the same as our previous work.. The primary antibodies Actin (20536‐1‐AP), LC3II (18725‐1‐AP), p62 (18420‐1‐AP) were purchased from Proteintech; TFEB (37785 S), pT389‐S6K (9234 S), p‐S6 (4858 S), S6K (9202 S), S6 (2217 S), p‐AMPKα (50081 S), AMPKα (5832 S), Raptor (2280 S) and TSC2 (D93F12) were purchased from Cell Signaling Technology; Flag (db7002) and HA (db2603) were purchased from Hangzhou Bio Technolog.. The secondary antibodies were obtained from Sigma.The secondary antibodies were obtained from Sigma.

    Article Title: A translational program that suppresses metabolism to shield the genome
    Article Snippet: Blots were stripped and reprobed when looking at total versus modified proteins unless otherwise noted.. Antibodies (all 1:1000): GLUT1 (Novus Biologicals, # NB110-39113), PAI-1 (Proteintech, #13801-1-AP), Puromycin (3RH11) (Kerafast, #EQ0001), β-ACTIN (C4) (SantaCruz Biotechnology, #sc-47778), EIF5A (abcam, #ab32443), EIF3D (Proteintech, # 10219-1-AP), EIF3K (Proteintech, # 10640-1-AP), acetyl-EIF5A (Lys47) (Boster Bio, #P01727), CYR61 (Proteintech, #26689-1-AP), SDC4 (R&D Systems, #P31431), SIRT1 (Proteintech, #13161-1-AP), SIRT2 (Proteintech, #19655-1-AP), NFKB2 (Proteintech, #10409-2-AP), 4E-BP (Cell Signaling, #9452 S), Phospho-4E-BP (Ser65) (Cell Signaling, #9451 S), TSC2 (D93F12) (Cell Signaling, #4308S), gamma H2AX (Novus, NB100-384), H2AX (Abcam, Ab11175), CRM-1 (Novus, NB100-79802), XPO-4 (Abcam, ab133237), eIF5A2 (Thermo, #PA5-30770), Phospho-RAPTOR (Ser792) (Cell Signaling, #89146S), RAPTOR (Cell Signaling, #2280), Phospho-Tuberin/TSC2 (Ser1387) (Cell Signaling, #5584S), NDRG1 (Abcam, #ab37897), and Anti-hypusine (mAbHpu24) antibody was kindly provided by Genentech.. The eIF5A1 antibodies and siRNAs used in this study were specific to eIF5A1 and not eIF5A2 as described by the manufacturer and validated in Supplementary Fig. .

    Article Title: Identification of mechanically regulated phosphorylation sites on tuberin (TSC2) that control mechanistic target of rapamycin (mTOR) signaling
    Article Snippet: Images were quantified using ImageJ software (U.S. National Institutes of Health, Bethesda, MD; http://rsb.info.nih.gov/nih-image/).Images were quantified using ImageJ software (U.S. National Institutes of Health, Bethesda, MD; http://rsb.info.nih.gov/nih-image/).. Antibodies - P-p70(389) (1A5) (#9206, used to probe Myc-tagged P-p70), P-p70(389) (#9234, used to probe endogenous P-p70), total p70 (49D7) (#2708), Anti-P-Akt substrate (RxRxxS*/T*) (23C8D2) for detection of PTSC2 (#10001), P-JNK(Thr183/Tyr185) (98F2) (#4671), total JNK (#9252), total mTOR (#2972), total Rictor (D16H9) (#9476), P-PKB (Ser473) (#4060), P-PKB (Thr308) (#9275S), total PKB (#9272), P-S6 (Ser240/4) (#5364), P-S6 (Ser235/6) (#2211), total S6 (#2217S), total TSC1 (D43E2) (#6935), total TSC2 (D93F12) (#4308) were purchased from Cell Signaling Technology (Danvers, MA, USA).. Anti-puromycin (12D10) (#MABE343) was purchased from EMD Millipore (Billerica, MA, USA).Anti-puromycin (12D10) (#MABE343) was purchased from EMD Millipore (Billerica, MA, USA).

    Article Title: Chronic signaling via the metabolic checkpoint kinase mTORC1 induces macrophage granuloma formation and marks sarcoidosis progression
    Article Snippet: HRP-conjugated secondary antibodies (Bethyl) 1:10000 were applied for 1 hour at room temperature in 4% low-fat milk and proteins were visualized using ECL substrate (Thermo Scientific).HRP-conjugated secondary antibodies (Bethyl) 1:10000 were applied for 1 hour at room temperature in 4% low-fat milk and proteins were visualized using ECL substrate (Thermo Scientific).. Primary antibodies were TSC2 (D93F12) 1:1000, p-Akt S473 (D9E) 1:1000, 4E-BP1 (53H11) 1:1000, p-S6 S240/244 (rabbit polyclonal) 1:3000, S6 (54D2) 1:1000, p-Akt T308 (D25E6) 1:1000, pan-Akt (40D4) 1:1000, β-tubulin (9F3) 1:1000 , p-RB Ser807/811 (D20B12) 1:1000, PDCD4 (D29C6) 1:1000, cleaved caspase 3 Asp175 (5A1E or rabbit polyclonal) 1:500, survivin (71G4B7) 1:1000 and p-S6K1 T389 (108D2) 1:1000, p-TSC2 S939 (rabbit polyclonal) 1:1000 obtained from Cell Signaling Technology.. Bcl-2 (N-19) 1:500, cyclin D1 (M-20) 1:500, p27Kip1 (C-19) 1:1000, cyclin A (C-19) 1:500, CDK4 (C-22) 1:1000, CDK6 (H-96) 1:500, and E2F1 (KH95) 1:500 were fom Santa Cruz. α-tubulin (CP06) 1:3000 was purchased from Calbiochem and GAPDH (rabbit polyclonal) 1:10000 from Trevigen.Bcl-2 (N-19) 1:500, cyclin D1 (M-20) 1:500, p27Kip1 (C-19) 1:1000, cyclin A (C-19) 1:500, CDK4 (C-22) 1:1000, CDK6 (H-96) 1:500, and E2F1 (KH95) 1:500 were fom Santa Cruz. α-tubulin (CP06) 1:3000 was purchased..

    Article Title: TSC1 promotes i NKT cell anergy and inhibits i NKT cell-mediated anti-tumor immunity
    Article Snippet: Fluorescence-conjugated anti-mouse TCRβ (H57-597), NK1.1 (PK136), CD44(IM7), B220(RA3-6B2), Gr-1(RB6-8C5), CD11c(N418), CD11b(M1/70), CD4 (GK1.5), CD8 (53-6.7), IFN-γ (XMG1.2), IL-4 (11B11), PD-1(RMP1-30), and ICOS(D10.G4.1) antibodies were purchased from BioLegend.Fluorescence-conjugated anti-mouse TCRβ (H57-597), NK1.1 (PK136), CD44(IM7), B220(RA3-6B2), Gr-1(RB6-8C5), CD11c(N418), CD11b(M1/70), CD4 (GK1.5), CD8 (53-6.7), IFN-γ (XMG1.2), IL-4 (11B11), PD-1(RMP1-30), and ICOS(D10.G4.1) antibodies were purchased from BioLegend.. Anti–phospho-p44/42 mitogen-activated protein kinase T202/Y204 (197G2), anti–phospho-S6 S240/244 (D68F8), anti–phospho-4E-BP1 T37/46 (236B4), anti–phospho-Akt S473 (193H12), TSC1 (D43E2), and TSC2 (D93F12) were purchased from Cell Signaling Technology, Inc. Anti–β-actin antibody was purchased from Sigma-Aldrich.. Cell surface staining was performed with 2% FBS-PBS.Cell surface staining was performed with 2% FBS-PBS.

    Article Title: TSC1 promotes i NKT cell anergy and inhibits i NKT cell-mediated anti-tumor immunity
    Article Snippet: CD11b(M1/70), CD4 (GK1.5), CD8 (53-6.7), IFN-γ (XMG1.2), IL-4 (11B11), PD-1(RMP1-30), and ICOS(D10.G4.1) antibodies were purchased from BioLegend. .. Anti–phospho-p44/42 mitogen-activated protein kinase T202/Y204 (197G2), anti–phospho-S6 S240/244 (D68F8), anti–phospho-4E-BP1 T37/46 (236B4), anti–phospho-Akt S473 (193H12), TSC1 (D43E2), and TSC2 (D93F12) were purchased from Cell Signaling Technology, Inc. Anti–β-actin antibody was purchased from Sigma-Aldrich. .. Cell surface staining was performed with 2% FBS-PBS.

    Labeling:

    Article Title: Trans‐gnetin H isolated from the seeds of Paeonia species induces autophagy via inhibiting mTORC1 signalling through AMPK activation
    Article Snippet: The WB protocol was the same as our previous work.The WB protocol was the same as our previous work.. The primary antibodies Actin (20536‐1‐AP), LC3II (18725‐1‐AP), p62 (18420‐1‐AP) were purchased from Proteintech; TFEB (37785 S), pT389‐S6K (9234 S), p‐S6 (4858 S), S6K (9202 S), S6 (2217 S), p‐AMPKα (50081 S), AMPKα (5832 S), Raptor (2280 S) and TSC2 (D93F12) were purchased from Cell Signaling Technology; Flag (db7002) and HA (db2603) were purchased from Hangzhou Bio Technolog.. The secondary antibodies were obtained from Sigma.The secondary antibodies were obtained from Sigma.

    Article Title: A translational program that suppresses metabolism to shield the genome
    Article Snippet: Blots were stripped and reprobed when looking at total versus modified proteins unless otherwise noted.. Antibodies (all 1:1000): GLUT1 (Novus Biologicals, # NB110-39113), PAI-1 (Proteintech, #13801-1-AP), Puromycin (3RH11) (Kerafast, #EQ0001), β-ACTIN (C4) (SantaCruz Biotechnology, #sc-47778), EIF5A (abcam, #ab32443), EIF3D (Proteintech, # 10219-1-AP), EIF3K (Proteintech, # 10640-1-AP), acetyl-EIF5A (Lys47) (Boster Bio, #P01727), CYR61 (Proteintech, #26689-1-AP), SDC4 (R&D Systems, #P31431), SIRT1 (Proteintech, #13161-1-AP), SIRT2 (Proteintech, #19655-1-AP), NFKB2 (Proteintech, #10409-2-AP), 4E-BP (Cell Signaling, #9452 S), Phospho-4E-BP (Ser65) (Cell Signaling, #9451 S), TSC2 (D93F12) (Cell Signaling, #4308S), gamma H2AX (Novus, NB100-384), H2AX (Abcam, Ab11175), CRM-1 (Novus, NB100-79802), XPO-4 (Abcam, ab133237), eIF5A2 (Thermo, #PA5-30770), Phospho-RAPTOR (Ser792) (Cell Signaling, #89146S), RAPTOR (Cell Signaling, #2280), Phospho-Tuberin/TSC2 (Ser1387) (Cell Signaling, #5584S), NDRG1 (Abcam, #ab37897), and Anti-hypusine (mAbHpu24) antibody was kindly provided by Genentech.. The eIF5A1 antibodies and siRNAs used in this study were specific to eIF5A1 and not eIF5A2 as described by the manufacturer and validated in Supplementary Fig. .

    Article Title: Identification of mechanically regulated phosphorylation sites on tuberin (TSC2) that control mechanistic target of rapamycin (mTOR) signaling
    Article Snippet: Images were quantified using ImageJ software (U.S. National Institutes of Health, Bethesda, MD; http://rsb.info.nih.gov/nih-image/).Images were quantified using ImageJ software (U.S. National Institutes of Health, Bethesda, MD; http://rsb.info.nih.gov/nih-image/).. Antibodies - P-p70(389) (1A5) (#9206, used to probe Myc-tagged P-p70), P-p70(389) (#9234, used to probe endogenous P-p70), total p70 (49D7) (#2708), Anti-P-Akt substrate (RxRxxS*/T*) (23C8D2) for detection of PTSC2 (#10001), P-JNK(Thr183/Tyr185) (98F2) (#4671), total JNK (#9252), total mTOR (#2972), total Rictor (D16H9) (#9476), P-PKB (Ser473) (#4060), P-PKB (Thr308) (#9275S), total PKB (#9272), P-S6 (Ser240/4) (#5364), P-S6 (Ser235/6) (#2211), total S6 (#2217S), total TSC1 (D43E2) (#6935), total TSC2 (D93F12) (#4308) were purchased from Cell Signaling Technology (Danvers, MA, USA).. Anti-puromycin (12D10) (#MABE343) was purchased from EMD Millipore (Billerica, MA, USA).Anti-puromycin (12D10) (#MABE343) was purchased from EMD Millipore (Billerica, MA, USA).

    Article Title: Chronic signaling via the metabolic checkpoint kinase mTORC1 induces macrophage granuloma formation and marks sarcoidosis progression
    Article Snippet: HRP-conjugated secondary antibodies (Bethyl) 1:10000 were applied for 1 hour at room temperature in 4% low-fat milk and proteins were visualized using ECL substrate (Thermo Scientific).HRP-conjugated secondary antibodies (Bethyl) 1:10000 were applied for 1 hour at room temperature in 4% low-fat milk and proteins were visualized using ECL substrate (Thermo Scientific).. Primary antibodies were TSC2 (D93F12) 1:1000, p-Akt S473 (D9E) 1:1000, 4E-BP1 (53H11) 1:1000, p-S6 S240/244 (rabbit polyclonal) 1:3000, S6 (54D2) 1:1000, p-Akt T308 (D25E6) 1:1000, pan-Akt (40D4) 1:1000, β-tubulin (9F3) 1:1000 , p-RB Ser807/811 (D20B12) 1:1000, PDCD4 (D29C6) 1:1000, cleaved caspase 3 Asp175 (5A1E or rabbit polyclonal) 1:500, survivin (71G4B7) 1:1000 and p-S6K1 T389 (108D2) 1:1000, p-TSC2 S939 (rabbit polyclonal) 1:1000 obtained from Cell Signaling Technology.. Bcl-2 (N-19) 1:500, cyclin D1 (M-20) 1:500, p27Kip1 (C-19) 1:1000, cyclin A (C-19) 1:500, CDK4 (C-22) 1:1000, CDK6 (H-96) 1:500, and E2F1 (KH95) 1:500 were fom Santa Cruz. α-tubulin (CP06) 1:3000 was purchased from Calbiochem and GAPDH (rabbit polyclonal) 1:10000 from Trevigen.Bcl-2 (N-19) 1:500, cyclin D1 (M-20) 1:500, p27Kip1 (C-19) 1:1000, cyclin A (C-19) 1:500, CDK4 (C-22) 1:1000, CDK6 (H-96) 1:500, and E2F1 (KH95) 1:500 were fom Santa Cruz. α-tubulin (CP06) 1:3000 was purchased..

    Article Title: TSC1 promotes i NKT cell anergy and inhibits i NKT cell-mediated anti-tumor immunity
    Article Snippet: Fluorescence-conjugated anti-mouse TCRβ (H57-597), NK1.1 (PK136), CD44(IM7), B220(RA3-6B2), Gr-1(RB6-8C5), CD11c(N418), CD11b(M1/70), CD4 (GK1.5), CD8 (53-6.7), IFN-γ (XMG1.2), IL-4 (11B11), PD-1(RMP1-30), and ICOS(D10.G4.1) antibodies were purchased from BioLegend.Fluorescence-conjugated anti-mouse TCRβ (H57-597), NK1.1 (PK136), CD44(IM7), B220(RA3-6B2), Gr-1(RB6-8C5), CD11c(N418), CD11b(M1/70), CD4 (GK1.5), CD8 (53-6.7), IFN-γ (XMG1.2), IL-4 (11B11), PD-1(RMP1-30), and ICOS(D10.G4.1) antibodies were purchased from BioLegend.. Anti–phospho-p44/42 mitogen-activated protein kinase T202/Y204 (197G2), anti–phospho-S6 S240/244 (D68F8), anti–phospho-4E-BP1 T37/46 (236B4), anti–phospho-Akt S473 (193H12), TSC1 (D43E2), and TSC2 (D93F12) were purchased from Cell Signaling Technology, Inc. Anti–β-actin antibody was purchased from Sigma-Aldrich.. Cell surface staining was performed with 2% FBS-PBS.Cell surface staining was performed with 2% FBS-PBS.

    Article Title: TSC1 promotes i NKT cell anergy and inhibits i NKT cell-mediated anti-tumor immunity
    Article Snippet: CD11b(M1/70), CD4 (GK1.5), CD8 (53-6.7), IFN-γ (XMG1.2), IL-4 (11B11), PD-1(RMP1-30), and ICOS(D10.G4.1) antibodies were purchased from BioLegend. .. Anti–phospho-p44/42 mitogen-activated protein kinase T202/Y204 (197G2), anti–phospho-S6 S240/244 (D68F8), anti–phospho-4E-BP1 T37/46 (236B4), anti–phospho-Akt S473 (193H12), TSC1 (D43E2), and TSC2 (D93F12) were purchased from Cell Signaling Technology, Inc. Anti–β-actin antibody was purchased from Sigma-Aldrich. .. Cell surface staining was performed with 2% FBS-PBS.

    Injection:

    Article Title: Trans‐gnetin H isolated from the seeds of Paeonia species induces autophagy via inhibiting mTORC1 signalling through AMPK activation
    Article Snippet: The WB protocol was the same as our previous work.The WB protocol was the same as our previous work.. The primary antibodies Actin (20536‐1‐AP), LC3II (18725‐1‐AP), p62 (18420‐1‐AP) were purchased from Proteintech; TFEB (37785 S), pT389‐S6K (9234 S), p‐S6 (4858 S), S6K (9202 S), S6 (2217 S), p‐AMPKα (50081 S), AMPKα (5832 S), Raptor (2280 S) and TSC2 (D93F12) were purchased from Cell Signaling Technology; Flag (db7002) and HA (db2603) were purchased from Hangzhou Bio Technolog.. The secondary antibodies were obtained from Sigma.The secondary antibodies were obtained from Sigma.

    Article Title: A translational program that suppresses metabolism to shield the genome
    Article Snippet: Blots were stripped and reprobed when looking at total versus modified proteins unless otherwise noted.. Antibodies (all 1:1000): GLUT1 (Novus Biologicals, # NB110-39113), PAI-1 (Proteintech, #13801-1-AP), Puromycin (3RH11) (Kerafast, #EQ0001), β-ACTIN (C4) (SantaCruz Biotechnology, #sc-47778), EIF5A (abcam, #ab32443), EIF3D (Proteintech, # 10219-1-AP), EIF3K (Proteintech, # 10640-1-AP), acetyl-EIF5A (Lys47) (Boster Bio, #P01727), CYR61 (Proteintech, #26689-1-AP), SDC4 (R&D Systems, #P31431), SIRT1 (Proteintech, #13161-1-AP), SIRT2 (Proteintech, #19655-1-AP), NFKB2 (Proteintech, #10409-2-AP), 4E-BP (Cell Signaling, #9452 S), Phospho-4E-BP (Ser65) (Cell Signaling, #9451 S), TSC2 (D93F12) (Cell Signaling, #4308S), gamma H2AX (Novus, NB100-384), H2AX (Abcam, Ab11175), CRM-1 (Novus, NB100-79802), XPO-4 (Abcam, ab133237), eIF5A2 (Thermo, #PA5-30770), Phospho-RAPTOR (Ser792) (Cell Signaling, #89146S), RAPTOR (Cell Signaling, #2280), Phospho-Tuberin/TSC2 (Ser1387) (Cell Signaling, #5584S), NDRG1 (Abcam, #ab37897), and Anti-hypusine (mAbHpu24) antibody was kindly provided by Genentech.. The eIF5A1 antibodies and siRNAs used in this study were specific to eIF5A1 and not eIF5A2 as described by the manufacturer and validated in Supplementary Fig. .

    Article Title: Identification of mechanically regulated phosphorylation sites on tuberin (TSC2) that control mechanistic target of rapamycin (mTOR) signaling
    Article Snippet: Images were quantified using ImageJ software (U.S. National Institutes of Health, Bethesda, MD; http://rsb.info.nih.gov/nih-image/).Images were quantified using ImageJ software (U.S. National Institutes of Health, Bethesda, MD; http://rsb.info.nih.gov/nih-image/).. Antibodies - P-p70(389) (1A5) (#9206, used to probe Myc-tagged P-p70), P-p70(389) (#9234, used to probe endogenous P-p70), total p70 (49D7) (#2708), Anti-P-Akt substrate (RxRxxS*/T*) (23C8D2) for detection of PTSC2 (#10001), P-JNK(Thr183/Tyr185) (98F2) (#4671), total JNK (#9252), total mTOR (#2972), total Rictor (D16H9) (#9476), P-PKB (Ser473) (#4060), P-PKB (Thr308) (#9275S), total PKB (#9272), P-S6 (Ser240/4) (#5364), P-S6 (Ser235/6) (#2211), total S6 (#2217S), total TSC1 (D43E2) (#6935), total TSC2 (D93F12) (#4308) were purchased from Cell Signaling Technology (Danvers, MA, USA).. Anti-puromycin (12D10) (#MABE343) was purchased from EMD Millipore (Billerica, MA, USA).Anti-puromycin (12D10) (#MABE343) was purchased from EMD Millipore (Billerica, MA, USA).

    Article Title: Chronic signaling via the metabolic checkpoint kinase mTORC1 induces macrophage granuloma formation and marks sarcoidosis progression
    Article Snippet: HRP-conjugated secondary antibodies (Bethyl) 1:10000 were applied for 1 hour at room temperature in 4% low-fat milk and proteins were visualized using ECL substrate (Thermo Scientific).HRP-conjugated secondary antibodies (Bethyl) 1:10000 were applied for 1 hour at room temperature in 4% low-fat milk and proteins were visualized using ECL substrate (Thermo Scientific).. Primary antibodies were TSC2 (D93F12) 1:1000, p-Akt S473 (D9E) 1:1000, 4E-BP1 (53H11) 1:1000, p-S6 S240/244 (rabbit polyclonal) 1:3000, S6 (54D2) 1:1000, p-Akt T308 (D25E6) 1:1000, pan-Akt (40D4) 1:1000, β-tubulin (9F3) 1:1000 , p-RB Ser807/811 (D20B12) 1:1000, PDCD4 (D29C6) 1:1000, cleaved caspase 3 Asp175 (5A1E or rabbit polyclonal) 1:500, survivin (71G4B7) 1:1000 and p-S6K1 T389 (108D2) 1:1000, p-TSC2 S939 (rabbit polyclonal) 1:1000 obtained from Cell Signaling Technology.. Bcl-2 (N-19) 1:500, cyclin D1 (M-20) 1:500, p27Kip1 (C-19) 1:1000, cyclin A (C-19) 1:500, CDK4 (C-22) 1:1000, CDK6 (H-96) 1:500, and E2F1 (KH95) 1:500 were fom Santa Cruz. α-tubulin (CP06) 1:3000 was purchased from Calbiochem and GAPDH (rabbit polyclonal) 1:10000 from Trevigen.Bcl-2 (N-19) 1:500, cyclin D1 (M-20) 1:500, p27Kip1 (C-19) 1:1000, cyclin A (C-19) 1:500, CDK4 (C-22) 1:1000, CDK6 (H-96) 1:500, and E2F1 (KH95) 1:500 were fom Santa Cruz. α-tubulin (CP06) 1:3000 was purchased..

    Article Title: TSC1 promotes i NKT cell anergy and inhibits i NKT cell-mediated anti-tumor immunity
    Article Snippet: Fluorescence-conjugated anti-mouse TCRβ (H57-597), NK1.1 (PK136), CD44(IM7), B220(RA3-6B2), Gr-1(RB6-8C5), CD11c(N418), CD11b(M1/70), CD4 (GK1.5), CD8 (53-6.7), IFN-γ (XMG1.2), IL-4 (11B11), PD-1(RMP1-30), and ICOS(D10.G4.1) antibodies were purchased from BioLegend.Fluorescence-conjugated anti-mouse TCRβ (H57-597), NK1.1 (PK136), CD44(IM7), B220(RA3-6B2), Gr-1(RB6-8C5), CD11c(N418), CD11b(M1/70), CD4 (GK1.5), CD8 (53-6.7), IFN-γ (XMG1.2), IL-4 (11B11), PD-1(RMP1-30), and ICOS(D10.G4.1) antibodies were purchased from BioLegend.. Anti–phospho-p44/42 mitogen-activated protein kinase T202/Y204 (197G2), anti–phospho-S6 S240/244 (D68F8), anti–phospho-4E-BP1 T37/46 (236B4), anti–phospho-Akt S473 (193H12), TSC1 (D43E2), and TSC2 (D93F12) were purchased from Cell Signaling Technology, Inc. Anti–β-actin antibody was purchased from Sigma-Aldrich.. Cell surface staining was performed with 2% FBS-PBS.Cell surface staining was performed with 2% FBS-PBS.

    Article Title: TSC1 promotes i NKT cell anergy and inhibits i NKT cell-mediated anti-tumor immunity
    Article Snippet: CD11b(M1/70), CD4 (GK1.5), CD8 (53-6.7), IFN-γ (XMG1.2), IL-4 (11B11), PD-1(RMP1-30), and ICOS(D10.G4.1) antibodies were purchased from BioLegend. .. Anti–phospho-p44/42 mitogen-activated protein kinase T202/Y204 (197G2), anti–phospho-S6 S240/244 (D68F8), anti–phospho-4E-BP1 T37/46 (236B4), anti–phospho-Akt S473 (193H12), TSC1 (D43E2), and TSC2 (D93F12) were purchased from Cell Signaling Technology, Inc. Anti–β-actin antibody was purchased from Sigma-Aldrich. .. Cell surface staining was performed with 2% FBS-PBS.

    Immunohistochemistry:

    Article Title: Trans‐gnetin H isolated from the seeds of Paeonia species induces autophagy via inhibiting mTORC1 signalling through AMPK activation
    Article Snippet: The WB protocol was the same as our previous work.The WB protocol was the same as our previous work.. The primary antibodies Actin (20536‐1‐AP), LC3II (18725‐1‐AP), p62 (18420‐1‐AP) were purchased from Proteintech; TFEB (37785 S), pT389‐S6K (9234 S), p‐S6 (4858 S), S6K (9202 S), S6 (2217 S), p‐AMPKα (50081 S), AMPKα (5832 S), Raptor (2280 S) and TSC2 (D93F12) were purchased from Cell Signaling Technology; Flag (db7002) and HA (db2603) were purchased from Hangzhou Bio Technolog.. The secondary antibodies were obtained from Sigma.The secondary antibodies were obtained from Sigma.

    Article Title: A translational program that suppresses metabolism to shield the genome
    Article Snippet: Blots were stripped and reprobed when looking at total versus modified proteins unless otherwise noted.. Antibodies (all 1:1000): GLUT1 (Novus Biologicals, # NB110-39113), PAI-1 (Proteintech, #13801-1-AP), Puromycin (3RH11) (Kerafast, #EQ0001), β-ACTIN (C4) (SantaCruz Biotechnology, #sc-47778), EIF5A (abcam, #ab32443), EIF3D (Proteintech, # 10219-1-AP), EIF3K (Proteintech, # 10640-1-AP), acetyl-EIF5A (Lys47) (Boster Bio, #P01727), CYR61 (Proteintech, #26689-1-AP), SDC4 (R&D Systems, #P31431), SIRT1 (Proteintech, #13161-1-AP), SIRT2 (Proteintech, #19655-1-AP), NFKB2 (Proteintech, #10409-2-AP), 4E-BP (Cell Signaling, #9452 S), Phospho-4E-BP (Ser65) (Cell Signaling, #9451 S), TSC2 (D93F12) (Cell Signaling, #4308S), gamma H2AX (Novus, NB100-384), H2AX (Abcam, Ab11175), CRM-1 (Novus, NB100-79802), XPO-4 (Abcam, ab133237), eIF5A2 (Thermo, #PA5-30770), Phospho-RAPTOR (Ser792) (Cell Signaling, #89146S), RAPTOR (Cell Signaling, #2280), Phospho-Tuberin/TSC2 (Ser1387) (Cell Signaling, #5584S), NDRG1 (Abcam, #ab37897), and Anti-hypusine (mAbHpu24) antibody was kindly provided by Genentech.. The eIF5A1 antibodies and siRNAs used in this study were specific to eIF5A1 and not eIF5A2 as described by the manufacturer and validated in Supplementary Fig. .

    Article Title: Identification of mechanically regulated phosphorylation sites on tuberin (TSC2) that control mechanistic target of rapamycin (mTOR) signaling
    Article Snippet: Images were quantified using ImageJ software (U.S. National Institutes of Health, Bethesda, MD; http://rsb.info.nih.gov/nih-image/).Images were quantified using ImageJ software (U.S. National Institutes of Health, Bethesda, MD; http://rsb.info.nih.gov/nih-image/).. Antibodies - P-p70(389) (1A5) (#9206, used to probe Myc-tagged P-p70), P-p70(389) (#9234, used to probe endogenous P-p70), total p70 (49D7) (#2708), Anti-P-Akt substrate (RxRxxS*/T*) (23C8D2) for detection of PTSC2 (#10001), P-JNK(Thr183/Tyr185) (98F2) (#4671), total JNK (#9252), total mTOR (#2972), total Rictor (D16H9) (#9476), P-PKB (Ser473) (#4060), P-PKB (Thr308) (#9275S), total PKB (#9272), P-S6 (Ser240/4) (#5364), P-S6 (Ser235/6) (#2211), total S6 (#2217S), total TSC1 (D43E2) (#6935), total TSC2 (D93F12) (#4308) were purchased from Cell Signaling Technology (Danvers, MA, USA).. Anti-puromycin (12D10) (#MABE343) was purchased from EMD Millipore (Billerica, MA, USA).Anti-puromycin (12D10) (#MABE343) was purchased from EMD Millipore (Billerica, MA, USA).

    Article Title: Chronic signaling via the metabolic checkpoint kinase mTORC1 induces macrophage granuloma formation and marks sarcoidosis progression
    Article Snippet: HRP-conjugated secondary antibodies (Bethyl) 1:10000 were applied for 1 hour at room temperature in 4% low-fat milk and proteins were visualized using ECL substrate (Thermo Scientific).HRP-conjugated secondary antibodies (Bethyl) 1:10000 were applied for 1 hour at room temperature in 4% low-fat milk and proteins were visualized using ECL substrate (Thermo Scientific).. Primary antibodies were TSC2 (D93F12) 1:1000, p-Akt S473 (D9E) 1:1000, 4E-BP1 (53H11) 1:1000, p-S6 S240/244 (rabbit polyclonal) 1:3000, S6 (54D2) 1:1000, p-Akt T308 (D25E6) 1:1000, pan-Akt (40D4) 1:1000, β-tubulin (9F3) 1:1000 , p-RB Ser807/811 (D20B12) 1:1000, PDCD4 (D29C6) 1:1000, cleaved caspase 3 Asp175 (5A1E or rabbit polyclonal) 1:500, survivin (71G4B7) 1:1000 and p-S6K1 T389 (108D2) 1:1000, p-TSC2 S939 (rabbit polyclonal) 1:1000 obtained from Cell Signaling Technology.. Bcl-2 (N-19) 1:500, cyclin D1 (M-20) 1:500, p27Kip1 (C-19) 1:1000, cyclin A (C-19) 1:500, CDK4 (C-22) 1:1000, CDK6 (H-96) 1:500, and E2F1 (KH95) 1:500 were fom Santa Cruz. α-tubulin (CP06) 1:3000 was purchased from Calbiochem and GAPDH (rabbit polyclonal) 1:10000 from Trevigen.Bcl-2 (N-19) 1:500, cyclin D1 (M-20) 1:500, p27Kip1 (C-19) 1:1000, cyclin A (C-19) 1:500, CDK4 (C-22) 1:1000, CDK6 (H-96) 1:500, and E2F1 (KH95) 1:500 were fom Santa Cruz. α-tubulin (CP06) 1:3000 was purchased..

    Article Title: TSC1 promotes i NKT cell anergy and inhibits i NKT cell-mediated anti-tumor immunity
    Article Snippet: Fluorescence-conjugated anti-mouse TCRβ (H57-597), NK1.1 (PK136), CD44(IM7), B220(RA3-6B2), Gr-1(RB6-8C5), CD11c(N418), CD11b(M1/70), CD4 (GK1.5), CD8 (53-6.7), IFN-γ (XMG1.2), IL-4 (11B11), PD-1(RMP1-30), and ICOS(D10.G4.1) antibodies were purchased from BioLegend.Fluorescence-conjugated anti-mouse TCRβ (H57-597), NK1.1 (PK136), CD44(IM7), B220(RA3-6B2), Gr-1(RB6-8C5), CD11c(N418), CD11b(M1/70), CD4 (GK1.5), CD8 (53-6.7), IFN-γ (XMG1.2), IL-4 (11B11), PD-1(RMP1-30), and ICOS(D10.G4.1) antibodies were purchased from BioLegend.. Anti–phospho-p44/42 mitogen-activated protein kinase T202/Y204 (197G2), anti–phospho-S6 S240/244 (D68F8), anti–phospho-4E-BP1 T37/46 (236B4), anti–phospho-Akt S473 (193H12), TSC1 (D43E2), and TSC2 (D93F12) were purchased from Cell Signaling Technology, Inc. Anti–β-actin antibody was purchased from Sigma-Aldrich.. Cell surface staining was performed with 2% FBS-PBS.Cell surface staining was performed with 2% FBS-PBS.

    Article Title: TSC1 promotes i NKT cell anergy and inhibits i NKT cell-mediated anti-tumor immunity
    Article Snippet: CD11b(M1/70), CD4 (GK1.5), CD8 (53-6.7), IFN-γ (XMG1.2), IL-4 (11B11), PD-1(RMP1-30), and ICOS(D10.G4.1) antibodies were purchased from BioLegend. .. Anti–phospho-p44/42 mitogen-activated protein kinase T202/Y204 (197G2), anti–phospho-S6 S240/244 (D68F8), anti–phospho-4E-BP1 T37/46 (236B4), anti–phospho-Akt S473 (193H12), TSC1 (D43E2), and TSC2 (D93F12) were purchased from Cell Signaling Technology, Inc. Anti–β-actin antibody was purchased from Sigma-Aldrich. .. Cell surface staining was performed with 2% FBS-PBS.

    Irradiation:

    Article Title: Trans‐gnetin H isolated from the seeds of Paeonia species induces autophagy via inhibiting mTORC1 signalling through AMPK activation
    Article Snippet: The WB protocol was the same as our previous work.The WB protocol was the same as our previous work.. The primary antibodies Actin (20536‐1‐AP), LC3II (18725‐1‐AP), p62 (18420‐1‐AP) were purchased from Proteintech; TFEB (37785 S), pT389‐S6K (9234 S), p‐S6 (4858 S), S6K (9202 S), S6 (2217 S), p‐AMPKα (50081 S), AMPKα (5832 S), Raptor (2280 S) and TSC2 (D93F12) were purchased from Cell Signaling Technology; Flag (db7002) and HA (db2603) were purchased from Hangzhou Bio Technolog.. The secondary antibodies were obtained from Sigma.The secondary antibodies were obtained from Sigma.

    Article Title: A translational program that suppresses metabolism to shield the genome
    Article Snippet: Blots were stripped and reprobed when looking at total versus modified proteins unless otherwise noted.. Antibodies (all 1:1000): GLUT1 (Novus Biologicals, # NB110-39113), PAI-1 (Proteintech, #13801-1-AP), Puromycin (3RH11) (Kerafast, #EQ0001), β-ACTIN (C4) (SantaCruz Biotechnology, #sc-47778), EIF5A (abcam, #ab32443), EIF3D (Proteintech, # 10219-1-AP), EIF3K (Proteintech, # 10640-1-AP), acetyl-EIF5A (Lys47) (Boster Bio, #P01727), CYR61 (Proteintech, #26689-1-AP), SDC4 (R&D Systems, #P31431), SIRT1 (Proteintech, #13161-1-AP), SIRT2 (Proteintech, #19655-1-AP), NFKB2 (Proteintech, #10409-2-AP), 4E-BP (Cell Signaling, #9452 S), Phospho-4E-BP (Ser65) (Cell Signaling, #9451 S), TSC2 (D93F12) (Cell Signaling, #4308S), gamma H2AX (Novus, NB100-384), H2AX (Abcam, Ab11175), CRM-1 (Novus, NB100-79802), XPO-4 (Abcam, ab133237), eIF5A2 (Thermo, #PA5-30770), Phospho-RAPTOR (Ser792) (Cell Signaling, #89146S), RAPTOR (Cell Signaling, #2280), Phospho-Tuberin/TSC2 (Ser1387) (Cell Signaling, #5584S), NDRG1 (Abcam, #ab37897), and Anti-hypusine (mAbHpu24) antibody was kindly provided by Genentech.. The eIF5A1 antibodies and siRNAs used in this study were specific to eIF5A1 and not eIF5A2 as described by the manufacturer and validated in Supplementary Fig. .

    Article Title: Identification of mechanically regulated phosphorylation sites on tuberin (TSC2) that control mechanistic target of rapamycin (mTOR) signaling
    Article Snippet: Images were quantified using ImageJ software (U.S. National Institutes of Health, Bethesda, MD; http://rsb.info.nih.gov/nih-image/).Images were quantified using ImageJ software (U.S. National Institutes of Health, Bethesda, MD; http://rsb.info.nih.gov/nih-image/).. Antibodies - P-p70(389) (1A5) (#9206, used to probe Myc-tagged P-p70), P-p70(389) (#9234, used to probe endogenous P-p70), total p70 (49D7) (#2708), Anti-P-Akt substrate (RxRxxS*/T*) (23C8D2) for detection of PTSC2 (#10001), P-JNK(Thr183/Tyr185) (98F2) (#4671), total JNK (#9252), total mTOR (#2972), total Rictor (D16H9) (#9476), P-PKB (Ser473) (#4060), P-PKB (Thr308) (#9275S), total PKB (#9272), P-S6 (Ser240/4) (#5364), P-S6 (Ser235/6) (#2211), total S6 (#2217S), total TSC1 (D43E2) (#6935), total TSC2 (D93F12) (#4308) were purchased from Cell Signaling Technology (Danvers, MA, USA).. Anti-puromycin (12D10) (#MABE343) was purchased from EMD Millipore (Billerica, MA, USA).Anti-puromycin (12D10) (#MABE343) was purchased from EMD Millipore (Billerica, MA, USA).

    Article Title: Chronic signaling via the metabolic checkpoint kinase mTORC1 induces macrophage granuloma formation and marks sarcoidosis progression
    Article Snippet: HRP-conjugated secondary antibodies (Bethyl) 1:10000 were applied for 1 hour at room temperature in 4% low-fat milk and proteins were visualized using ECL substrate (Thermo Scientific).HRP-conjugated secondary antibodies (Bethyl) 1:10000 were applied for 1 hour at room temperature in 4% low-fat milk and proteins were visualized using ECL substrate (Thermo Scientific).. Primary antibodies were TSC2 (D93F12) 1:1000, p-Akt S473 (D9E) 1:1000, 4E-BP1 (53H11) 1:1000, p-S6 S240/244 (rabbit polyclonal) 1:3000, S6 (54D2) 1:1000, p-Akt T308 (D25E6) 1:1000, pan-Akt (40D4) 1:1000, β-tubulin (9F3) 1:1000 , p-RB Ser807/811 (D20B12) 1:1000, PDCD4 (D29C6) 1:1000, cleaved caspase 3 Asp175 (5A1E or rabbit polyclonal) 1:500, survivin (71G4B7) 1:1000 and p-S6K1 T389 (108D2) 1:1000, p-TSC2 S939 (rabbit polyclonal) 1:1000 obtained from Cell Signaling Technology.. Bcl-2 (N-19) 1:500, cyclin D1 (M-20) 1:500, p27Kip1 (C-19) 1:1000, cyclin A (C-19) 1:500, CDK4 (C-22) 1:1000, CDK6 (H-96) 1:500, and E2F1 (KH95) 1:500 were fom Santa Cruz. α-tubulin (CP06) 1:3000 was purchased from Calbiochem and GAPDH (rabbit polyclonal) 1:10000 from Trevigen.Bcl-2 (N-19) 1:500, cyclin D1 (M-20) 1:500, p27Kip1 (C-19) 1:1000, cyclin A (C-19) 1:500, CDK4 (C-22) 1:1000, CDK6 (H-96) 1:500, and E2F1 (KH95) 1:500 were fom Santa Cruz. α-tubulin (CP06) 1:3000 was purchased..

    Article Title: TSC1 promotes i NKT cell anergy and inhibits i NKT cell-mediated anti-tumor immunity
    Article Snippet: Fluorescence-conjugated anti-mouse TCRβ (H57-597), NK1.1 (PK136), CD44(IM7), B220(RA3-6B2), Gr-1(RB6-8C5), CD11c(N418), CD11b(M1/70), CD4 (GK1.5), CD8 (53-6.7), IFN-γ (XMG1.2), IL-4 (11B11), PD-1(RMP1-30), and ICOS(D10.G4.1) antibodies were purchased from BioLegend.Fluorescence-conjugated anti-mouse TCRβ (H57-597), NK1.1 (PK136), CD44(IM7), B220(RA3-6B2), Gr-1(RB6-8C5), CD11c(N418), CD11b(M1/70), CD4 (GK1.5), CD8 (53-6.7), IFN-γ (XMG1.2), IL-4 (11B11), PD-1(RMP1-30), and ICOS(D10.G4.1) antibodies were purchased from BioLegend.. Anti–phospho-p44/42 mitogen-activated protein kinase T202/Y204 (197G2), anti–phospho-S6 S240/244 (D68F8), anti–phospho-4E-BP1 T37/46 (236B4), anti–phospho-Akt S473 (193H12), TSC1 (D43E2), and TSC2 (D93F12) were purchased from Cell Signaling Technology, Inc. Anti–β-actin antibody was purchased from Sigma-Aldrich.. Cell surface staining was performed with 2% FBS-PBS.Cell surface staining was performed with 2% FBS-PBS.

    Article Title: TSC1 promotes i NKT cell anergy and inhibits i NKT cell-mediated anti-tumor immunity
    Article Snippet: CD11b(M1/70), CD4 (GK1.5), CD8 (53-6.7), IFN-γ (XMG1.2), IL-4 (11B11), PD-1(RMP1-30), and ICOS(D10.G4.1) antibodies were purchased from BioLegend. .. Anti–phospho-p44/42 mitogen-activated protein kinase T202/Y204 (197G2), anti–phospho-S6 S240/244 (D68F8), anti–phospho-4E-BP1 T37/46 (236B4), anti–phospho-Akt S473 (193H12), TSC1 (D43E2), and TSC2 (D93F12) were purchased from Cell Signaling Technology, Inc. Anti–β-actin antibody was purchased from Sigma-Aldrich. .. Cell surface staining was performed with 2% FBS-PBS.

    Flow Cytometry:

    Article Title: Trans‐gnetin H isolated from the seeds of Paeonia species induces autophagy via inhibiting mTORC1 signalling through AMPK activation
    Article Snippet: The WB protocol was the same as our previous work.The WB protocol was the same as our previous work.. The primary antibodies Actin (20536‐1‐AP), LC3II (18725‐1‐AP), p62 (18420‐1‐AP) were purchased from Proteintech; TFEB (37785 S), pT389‐S6K (9234 S), p‐S6 (4858 S), S6K (9202 S), S6 (2217 S), p‐AMPKα (50081 S), AMPKα (5832 S), Raptor (2280 S) and TSC2 (D93F12) were purchased from Cell Signaling Technology; Flag (db7002) and HA (db2603) were purchased from Hangzhou Bio Technolog.. The secondary antibodies were obtained from Sigma.The secondary antibodies were obtained from Sigma.

    Article Title: A translational program that suppresses metabolism to shield the genome
    Article Snippet: Blots were stripped and reprobed when looking at total versus modified proteins unless otherwise noted.. Antibodies (all 1:1000): GLUT1 (Novus Biologicals, # NB110-39113), PAI-1 (Proteintech, #13801-1-AP), Puromycin (3RH11) (Kerafast, #EQ0001), β-ACTIN (C4) (SantaCruz Biotechnology, #sc-47778), EIF5A (abcam, #ab32443), EIF3D (Proteintech, # 10219-1-AP), EIF3K (Proteintech, # 10640-1-AP), acetyl-EIF5A (Lys47) (Boster Bio, #P01727), CYR61 (Proteintech, #26689-1-AP), SDC4 (R&D Systems, #P31431), SIRT1 (Proteintech, #13161-1-AP), SIRT2 (Proteintech, #19655-1-AP), NFKB2 (Proteintech, #10409-2-AP), 4E-BP (Cell Signaling, #9452 S), Phospho-4E-BP (Ser65) (Cell Signaling, #9451 S), TSC2 (D93F12) (Cell Signaling, #4308S), gamma H2AX (Novus, NB100-384), H2AX (Abcam, Ab11175), CRM-1 (Novus, NB100-79802), XPO-4 (Abcam, ab133237), eIF5A2 (Thermo, #PA5-30770), Phospho-RAPTOR (Ser792) (Cell Signaling, #89146S), RAPTOR (Cell Signaling, #2280), Phospho-Tuberin/TSC2 (Ser1387) (Cell Signaling, #5584S), NDRG1 (Abcam, #ab37897), and Anti-hypusine (mAbHpu24) antibody was kindly provided by Genentech.. The eIF5A1 antibodies and siRNAs used in this study were specific to eIF5A1 and not eIF5A2 as described by the manufacturer and validated in Supplementary Fig. .

    Article Title: Identification of mechanically regulated phosphorylation sites on tuberin (TSC2) that control mechanistic target of rapamycin (mTOR) signaling
    Article Snippet: Images were quantified using ImageJ software (U.S. National Institutes of Health, Bethesda, MD; http://rsb.info.nih.gov/nih-image/).Images were quantified using ImageJ software (U.S. National Institutes of Health, Bethesda, MD; http://rsb.info.nih.gov/nih-image/).. Antibodies - P-p70(389) (1A5) (#9206, used to probe Myc-tagged P-p70), P-p70(389) (#9234, used to probe endogenous P-p70), total p70 (49D7) (#2708), Anti-P-Akt substrate (RxRxxS*/T*) (23C8D2) for detection of PTSC2 (#10001), P-JNK(Thr183/Tyr185) (98F2) (#4671), total JNK (#9252), total mTOR (#2972), total Rictor (D16H9) (#9476), P-PKB (Ser473) (#4060), P-PKB (Thr308) (#9275S), total PKB (#9272), P-S6 (Ser240/4) (#5364), P-S6 (Ser235/6) (#2211), total S6 (#2217S), total TSC1 (D43E2) (#6935), total TSC2 (D93F12) (#4308) were purchased from Cell Signaling Technology (Danvers, MA, USA).. Anti-puromycin (12D10) (#MABE343) was purchased from EMD Millipore (Billerica, MA, USA).Anti-puromycin (12D10) (#MABE343) was purchased from EMD Millipore (Billerica, MA, USA).

    Article Title: Chronic signaling via the metabolic checkpoint kinase mTORC1 induces macrophage granuloma formation and marks sarcoidosis progression
    Article Snippet: HRP-conjugated secondary antibodies (Bethyl) 1:10000 were applied for 1 hour at room temperature in 4% low-fat milk and proteins were visualized using ECL substrate (Thermo Scientific).HRP-conjugated secondary antibodies (Bethyl) 1:10000 were applied for 1 hour at room temperature in 4% low-fat milk and proteins were visualized using ECL substrate (Thermo Scientific).. Primary antibodies were TSC2 (D93F12) 1:1000, p-Akt S473 (D9E) 1:1000, 4E-BP1 (53H11) 1:1000, p-S6 S240/244 (rabbit polyclonal) 1:3000, S6 (54D2) 1:1000, p-Akt T308 (D25E6) 1:1000, pan-Akt (40D4) 1:1000, β-tubulin (9F3) 1:1000 , p-RB Ser807/811 (D20B12) 1:1000, PDCD4 (D29C6) 1:1000, cleaved caspase 3 Asp175 (5A1E or rabbit polyclonal) 1:500, survivin (71G4B7) 1:1000 and p-S6K1 T389 (108D2) 1:1000, p-TSC2 S939 (rabbit polyclonal) 1:1000 obtained from Cell Signaling Technology.. Bcl-2 (N-19) 1:500, cyclin D1 (M-20) 1:500, p27Kip1 (C-19) 1:1000, cyclin A (C-19) 1:500, CDK4 (C-22) 1:1000, CDK6 (H-96) 1:500, and E2F1 (KH95) 1:500 were fom Santa Cruz. α-tubulin (CP06) 1:3000 was purchased from Calbiochem and GAPDH (rabbit polyclonal) 1:10000 from Trevigen.Bcl-2 (N-19) 1:500, cyclin D1 (M-20) 1:500, p27Kip1 (C-19) 1:1000, cyclin A (C-19) 1:500, CDK4 (C-22) 1:1000, CDK6 (H-96) 1:500, and E2F1 (KH95) 1:500 were fom Santa Cruz. α-tubulin (CP06) 1:3000 was purchased..

    Article Title: TSC1 promotes i NKT cell anergy and inhibits i NKT cell-mediated anti-tumor immunity
    Article Snippet: Fluorescence-conjugated anti-mouse TCRβ (H57-597), NK1.1 (PK136), CD44(IM7), B220(RA3-6B2), Gr-1(RB6-8C5), CD11c(N418), CD11b(M1/70), CD4 (GK1.5), CD8 (53-6.7), IFN-γ (XMG1.2), IL-4 (11B11), PD-1(RMP1-30), and ICOS(D10.G4.1) antibodies were purchased from BioLegend.Fluorescence-conjugated anti-mouse TCRβ (H57-597), NK1.1 (PK136), CD44(IM7), B220(RA3-6B2), Gr-1(RB6-8C5), CD11c(N418), CD11b(M1/70), CD4 (GK1.5), CD8 (53-6.7), IFN-γ (XMG1.2), IL-4 (11B11), PD-1(RMP1-30), and ICOS(D10.G4.1) antibodies were purchased from BioLegend.. Anti–phospho-p44/42 mitogen-activated protein kinase T202/Y204 (197G2), anti–phospho-S6 S240/244 (D68F8), anti–phospho-4E-BP1 T37/46 (236B4), anti–phospho-Akt S473 (193H12), TSC1 (D43E2), and TSC2 (D93F12) were purchased from Cell Signaling Technology, Inc. Anti–β-actin antibody was purchased from Sigma-Aldrich.. Cell surface staining was performed with 2% FBS-PBS.Cell surface staining was performed with 2% FBS-PBS.

    Article Title: TSC1 promotes i NKT cell anergy and inhibits i NKT cell-mediated anti-tumor immunity
    Article Snippet: CD11b(M1/70), CD4 (GK1.5), CD8 (53-6.7), IFN-γ (XMG1.2), IL-4 (11B11), PD-1(RMP1-30), and ICOS(D10.G4.1) antibodies were purchased from BioLegend. .. Anti–phospho-p44/42 mitogen-activated protein kinase T202/Y204 (197G2), anti–phospho-S6 S240/244 (D68F8), anti–phospho-4E-BP1 T37/46 (236B4), anti–phospho-Akt S473 (193H12), TSC1 (D43E2), and TSC2 (D93F12) were purchased from Cell Signaling Technology, Inc. Anti–β-actin antibody was purchased from Sigma-Aldrich. .. Cell surface staining was performed with 2% FBS-PBS.

    Derivative Assay:

    Article Title: Trans‐gnetin H isolated from the seeds of Paeonia species induces autophagy via inhibiting mTORC1 signalling through AMPK activation
    Article Snippet: The WB protocol was the same as our previous work.The WB protocol was the same as our previous work.. The primary antibodies Actin (20536‐1‐AP), LC3II (18725‐1‐AP), p62 (18420‐1‐AP) were purchased from Proteintech; TFEB (37785 S), pT389‐S6K (9234 S), p‐S6 (4858 S), S6K (9202 S), S6 (2217 S), p‐AMPKα (50081 S), AMPKα (5832 S), Raptor (2280 S) and TSC2 (D93F12) were purchased from Cell Signaling Technology; Flag (db7002) and HA (db2603) were purchased from Hangzhou Bio Technolog.. The secondary antibodies were obtained from Sigma.The secondary antibodies were obtained from Sigma.

    Article Title: A translational program that suppresses metabolism to shield the genome
    Article Snippet: Blots were stripped and reprobed when looking at total versus modified proteins unless otherwise noted.. Antibodies (all 1:1000): GLUT1 (Novus Biologicals, # NB110-39113), PAI-1 (Proteintech, #13801-1-AP), Puromycin (3RH11) (Kerafast, #EQ0001), β-ACTIN (C4) (SantaCruz Biotechnology, #sc-47778), EIF5A (abcam, #ab32443), EIF3D (Proteintech, # 10219-1-AP), EIF3K (Proteintech, # 10640-1-AP), acetyl-EIF5A (Lys47) (Boster Bio, #P01727), CYR61 (Proteintech, #26689-1-AP), SDC4 (R&D Systems, #P31431), SIRT1 (Proteintech, #13161-1-AP), SIRT2 (Proteintech, #19655-1-AP), NFKB2 (Proteintech, #10409-2-AP), 4E-BP (Cell Signaling, #9452 S), Phospho-4E-BP (Ser65) (Cell Signaling, #9451 S), TSC2 (D93F12) (Cell Signaling, #4308S), gamma H2AX (Novus, NB100-384), H2AX (Abcam, Ab11175), CRM-1 (Novus, NB100-79802), XPO-4 (Abcam, ab133237), eIF5A2 (Thermo, #PA5-30770), Phospho-RAPTOR (Ser792) (Cell Signaling, #89146S), RAPTOR (Cell Signaling, #2280), Phospho-Tuberin/TSC2 (Ser1387) (Cell Signaling, #5584S), NDRG1 (Abcam, #ab37897), and Anti-hypusine (mAbHpu24) antibody was kindly provided by Genentech.. The eIF5A1 antibodies and siRNAs used in this study were specific to eIF5A1 and not eIF5A2 as described by the manufacturer and validated in Supplementary Fig. .

    Article Title: Identification of mechanically regulated phosphorylation sites on tuberin (TSC2) that control mechanistic target of rapamycin (mTOR) signaling
    Article Snippet: Images were quantified using ImageJ software (U.S. National Institutes of Health, Bethesda, MD; http://rsb.info.nih.gov/nih-image/).Images were quantified using ImageJ software (U.S. National Institutes of Health, Bethesda, MD; http://rsb.info.nih.gov/nih-image/).. Antibodies - P-p70(389) (1A5) (#9206, used to probe Myc-tagged P-p70), P-p70(389) (#9234, used to probe endogenous P-p70), total p70 (49D7) (#2708), Anti-P-Akt substrate (RxRxxS*/T*) (23C8D2) for detection of PTSC2 (#10001), P-JNK(Thr183/Tyr185) (98F2) (#4671), total JNK (#9252), total mTOR (#2972), total Rictor (D16H9) (#9476), P-PKB (Ser473) (#4060), P-PKB (Thr308) (#9275S), total PKB (#9272), P-S6 (Ser240/4) (#5364), P-S6 (Ser235/6) (#2211), total S6 (#2217S), total TSC1 (D43E2) (#6935), total TSC2 (D93F12) (#4308) were purchased from Cell Signaling Technology (Danvers, MA, USA).. Anti-puromycin (12D10) (#MABE343) was purchased from EMD Millipore (Billerica, MA, USA).Anti-puromycin (12D10) (#MABE343) was purchased from EMD Millipore (Billerica, MA, USA).

    Article Title: Chronic signaling via the metabolic checkpoint kinase mTORC1 induces macrophage granuloma formation and marks sarcoidosis progression
    Article Snippet: HRP-conjugated secondary antibodies (Bethyl) 1:10000 were applied for 1 hour at room temperature in 4% low-fat milk and proteins were visualized using ECL substrate (Thermo Scientific).HRP-conjugated secondary antibodies (Bethyl) 1:10000 were applied for 1 hour at room temperature in 4% low-fat milk and proteins were visualized using ECL substrate (Thermo Scientific).. Primary antibodies were TSC2 (D93F12) 1:1000, p-Akt S473 (D9E) 1:1000, 4E-BP1 (53H11) 1:1000, p-S6 S240/244 (rabbit polyclonal) 1:3000, S6 (54D2) 1:1000, p-Akt T308 (D25E6) 1:1000, pan-Akt (40D4) 1:1000, β-tubulin (9F3) 1:1000 , p-RB Ser807/811 (D20B12) 1:1000, PDCD4 (D29C6) 1:1000, cleaved caspase 3 Asp175 (5A1E or rabbit polyclonal) 1:500, survivin (71G4B7) 1:1000 and p-S6K1 T389 (108D2) 1:1000, p-TSC2 S939 (rabbit polyclonal) 1:1000 obtained from Cell Signaling Technology.. Bcl-2 (N-19) 1:500, cyclin D1 (M-20) 1:500, p27Kip1 (C-19) 1:1000, cyclin A (C-19) 1:500, CDK4 (C-22) 1:1000, CDK6 (H-96) 1:500, and E2F1 (KH95) 1:500 were fom Santa Cruz. α-tubulin (CP06) 1:3000 was purchased from Calbiochem and GAPDH (rabbit polyclonal) 1:10000 from Trevigen.Bcl-2 (N-19) 1:500, cyclin D1 (M-20) 1:500, p27Kip1 (C-19) 1:1000, cyclin A (C-19) 1:500, CDK4 (C-22) 1:1000, CDK6 (H-96) 1:500, and E2F1 (KH95) 1:500 were fom Santa Cruz. α-tubulin (CP06) 1:3000 was purchased..

    Article Title: TSC1 promotes i NKT cell anergy and inhibits i NKT cell-mediated anti-tumor immunity
    Article Snippet: Fluorescence-conjugated anti-mouse TCRβ (H57-597), NK1.1 (PK136), CD44(IM7), B220(RA3-6B2), Gr-1(RB6-8C5), CD11c(N418), CD11b(M1/70), CD4 (GK1.5), CD8 (53-6.7), IFN-γ (XMG1.2), IL-4 (11B11), PD-1(RMP1-30), and ICOS(D10.G4.1) antibodies were purchased from BioLegend.Fluorescence-conjugated anti-mouse TCRβ (H57-597), NK1.1 (PK136), CD44(IM7), B220(RA3-6B2), Gr-1(RB6-8C5), CD11c(N418), CD11b(M1/70), CD4 (GK1.5), CD8 (53-6.7), IFN-γ (XMG1.2), IL-4 (11B11), PD-1(RMP1-30), and ICOS(D10.G4.1) antibodies were purchased from BioLegend.. Anti–phospho-p44/42 mitogen-activated protein kinase T202/Y204 (197G2), anti–phospho-S6 S240/244 (D68F8), anti–phospho-4E-BP1 T37/46 (236B4), anti–phospho-Akt S473 (193H12), TSC1 (D43E2), and TSC2 (D93F12) were purchased from Cell Signaling Technology, Inc. Anti–β-actin antibody was purchased from Sigma-Aldrich.. Cell surface staining was performed with 2% FBS-PBS.Cell surface staining was performed with 2% FBS-PBS.

    Article Title: TSC1 promotes i NKT cell anergy and inhibits i NKT cell-mediated anti-tumor immunity
    Article Snippet: CD11b(M1/70), CD4 (GK1.5), CD8 (53-6.7), IFN-γ (XMG1.2), IL-4 (11B11), PD-1(RMP1-30), and ICOS(D10.G4.1) antibodies were purchased from BioLegend. .. Anti–phospho-p44/42 mitogen-activated protein kinase T202/Y204 (197G2), anti–phospho-S6 S240/244 (D68F8), anti–phospho-4E-BP1 T37/46 (236B4), anti–phospho-Akt S473 (193H12), TSC1 (D43E2), and TSC2 (D93F12) were purchased from Cell Signaling Technology, Inc. Anti–β-actin antibody was purchased from Sigma-Aldrich. .. Cell surface staining was performed with 2% FBS-PBS.

    Solvent:

    Article Title: Trans‐gnetin H isolated from the seeds of Paeonia species induces autophagy via inhibiting mTORC1 signalling through AMPK activation
    Article Snippet: The WB protocol was the same as our previous work.The WB protocol was the same as our previous work.. The primary antibodies Actin (20536‐1‐AP), LC3II (18725‐1‐AP), p62 (18420‐1‐AP) were purchased from Proteintech; TFEB (37785 S), pT389‐S6K (9234 S), p‐S6 (4858 S), S6K (9202 S), S6 (2217 S), p‐AMPKα (50081 S), AMPKα (5832 S), Raptor (2280 S) and TSC2 (D93F12) were purchased from Cell Signaling Technology; Flag (db7002) and HA (db2603) were purchased from Hangzhou Bio Technolog.. The secondary antibodies were obtained from Sigma.The secondary antibodies were obtained from Sigma.

    Article Title: A translational program that suppresses metabolism to shield the genome
    Article Snippet: Blots were stripped and reprobed when looking at total versus modified proteins unless otherwise noted.. Antibodies (all 1:1000): GLUT1 (Novus Biologicals, # NB110-39113), PAI-1 (Proteintech, #13801-1-AP), Puromycin (3RH11) (Kerafast, #EQ0001), β-ACTIN (C4) (SantaCruz Biotechnology, #sc-47778), EIF5A (abcam, #ab32443), EIF3D (Proteintech, # 10219-1-AP), EIF3K (Proteintech, # 10640-1-AP), acetyl-EIF5A (Lys47) (Boster Bio, #P01727), CYR61 (Proteintech, #26689-1-AP), SDC4 (R&D Systems, #P31431), SIRT1 (Proteintech, #13161-1-AP), SIRT2 (Proteintech, #19655-1-AP), NFKB2 (Proteintech, #10409-2-AP), 4E-BP (Cell Signaling, #9452 S), Phospho-4E-BP (Ser65) (Cell Signaling, #9451 S), TSC2 (D93F12) (Cell Signaling, #4308S), gamma H2AX (Novus, NB100-384), H2AX (Abcam, Ab11175), CRM-1 (Novus, NB100-79802), XPO-4 (Abcam, ab133237), eIF5A2 (Thermo, #PA5-30770), Phospho-RAPTOR (Ser792) (Cell Signaling, #89146S), RAPTOR (Cell Signaling, #2280), Phospho-Tuberin/TSC2 (Ser1387) (Cell Signaling, #5584S), NDRG1 (Abcam, #ab37897), and Anti-hypusine (mAbHpu24) antibody was kindly provided by Genentech.. The eIF5A1 antibodies and siRNAs used in this study were specific to eIF5A1 and not eIF5A2 as described by the manufacturer and validated in Supplementary Fig. .

    Article Title: Identification of mechanically regulated phosphorylation sites on tuberin (TSC2) that control mechanistic target of rapamycin (mTOR) signaling
    Article Snippet: Images were quantified using ImageJ software (U.S. National Institutes of Health, Bethesda, MD; http://rsb.info.nih.gov/nih-image/).Images were quantified using ImageJ software (U.S. National Institutes of Health, Bethesda, MD; http://rsb.info.nih.gov/nih-image/).. Antibodies - P-p70(389) (1A5) (#9206, used to probe Myc-tagged P-p70), P-p70(389) (#9234, used to probe endogenous P-p70), total p70 (49D7) (#2708), Anti-P-Akt substrate (RxRxxS*/T*) (23C8D2) for detection of PTSC2 (#10001), P-JNK(Thr183/Tyr185) (98F2) (#4671), total JNK (#9252), total mTOR (#2972), total Rictor (D16H9) (#9476), P-PKB (Ser473) (#4060), P-PKB (Thr308) (#9275S), total PKB (#9272), P-S6 (Ser240/4) (#5364), P-S6 (Ser235/6) (#2211), total S6 (#2217S), total TSC1 (D43E2) (#6935), total TSC2 (D93F12) (#4308) were purchased from Cell Signaling Technology (Danvers, MA, USA).. Anti-puromycin (12D10) (#MABE343) was purchased from EMD Millipore (Billerica, MA, USA).Anti-puromycin (12D10) (#MABE343) was purchased from EMD Millipore (Billerica, MA, USA).

    Article Title: Chronic signaling via the metabolic checkpoint kinase mTORC1 induces macrophage granuloma formation and marks sarcoidosis progression
    Article Snippet: HRP-conjugated secondary antibodies (Bethyl) 1:10000 were applied for 1 hour at room temperature in 4% low-fat milk and proteins were visualized using ECL substrate (Thermo Scientific).HRP-conjugated secondary antibodies (Bethyl) 1:10000 were applied for 1 hour at room temperature in 4% low-fat milk and proteins were visualized using ECL substrate (Thermo Scientific).. Primary antibodies were TSC2 (D93F12) 1:1000, p-Akt S473 (D9E) 1:1000, 4E-BP1 (53H11) 1:1000, p-S6 S240/244 (rabbit polyclonal) 1:3000, S6 (54D2) 1:1000, p-Akt T308 (D25E6) 1:1000, pan-Akt (40D4) 1:1000, β-tubulin (9F3) 1:1000 , p-RB Ser807/811 (D20B12) 1:1000, PDCD4 (D29C6) 1:1000, cleaved caspase 3 Asp175 (5A1E or rabbit polyclonal) 1:500, survivin (71G4B7) 1:1000 and p-S6K1 T389 (108D2) 1:1000, p-TSC2 S939 (rabbit polyclonal) 1:1000 obtained from Cell Signaling Technology.. Bcl-2 (N-19) 1:500, cyclin D1 (M-20) 1:500, p27Kip1 (C-19) 1:1000, cyclin A (C-19) 1:500, CDK4 (C-22) 1:1000, CDK6 (H-96) 1:500, and E2F1 (KH95) 1:500 were fom Santa Cruz. α-tubulin (CP06) 1:3000 was purchased from Calbiochem and GAPDH (rabbit polyclonal) 1:10000 from Trevigen.Bcl-2 (N-19) 1:500, cyclin D1 (M-20) 1:500, p27Kip1 (C-19) 1:1000, cyclin A (C-19) 1:500, CDK4 (C-22) 1:1000, CDK6 (H-96) 1:500, and E2F1 (KH95) 1:500 were fom Santa Cruz. α-tubulin (CP06) 1:3000 was purchased..

    Article Title: TSC1 promotes i NKT cell anergy and inhibits i NKT cell-mediated anti-tumor immunity
    Article Snippet: Fluorescence-conjugated anti-mouse TCRβ (H57-597), NK1.1 (PK136), CD44(IM7), B220(RA3-6B2), Gr-1(RB6-8C5), CD11c(N418), CD11b(M1/70), CD4 (GK1.5), CD8 (53-6.7), IFN-γ (XMG1.2), IL-4 (11B11), PD-1(RMP1-30), and ICOS(D10.G4.1) antibodies were purchased from BioLegend.Fluorescence-conjugated anti-mouse TCRβ (H57-597), NK1.1 (PK136), CD44(IM7), B220(RA3-6B2), Gr-1(RB6-8C5), CD11c(N418), CD11b(M1/70), CD4 (GK1.5), CD8 (53-6.7), IFN-γ (XMG1.2), IL-4 (11B11), PD-1(RMP1-30), and ICOS(D10.G4.1) antibodies were purchased from BioLegend.. Anti–phospho-p44/42 mitogen-activated protein kinase T202/Y204 (197G2), anti–phospho-S6 S240/244 (D68F8), anti–phospho-4E-BP1 T37/46 (236B4), anti–phospho-Akt S473 (193H12), TSC1 (D43E2), and TSC2 (D93F12) were purchased from Cell Signaling Technology, Inc. Anti–β-actin antibody was purchased from Sigma-Aldrich.. Cell surface staining was performed with 2% FBS-PBS.Cell surface staining was performed with 2% FBS-PBS.

    Article Title: TSC1 promotes i NKT cell anergy and inhibits i NKT cell-mediated anti-tumor immunity
    Article Snippet: CD11b(M1/70), CD4 (GK1.5), CD8 (53-6.7), IFN-γ (XMG1.2), IL-4 (11B11), PD-1(RMP1-30), and ICOS(D10.G4.1) antibodies were purchased from BioLegend. .. Anti–phospho-p44/42 mitogen-activated protein kinase T202/Y204 (197G2), anti–phospho-S6 S240/244 (D68F8), anti–phospho-4E-BP1 T37/46 (236B4), anti–phospho-Akt S473 (193H12), TSC1 (D43E2), and TSC2 (D93F12) were purchased from Cell Signaling Technology, Inc. Anti–β-actin antibody was purchased from Sigma-Aldrich. .. Cell surface staining was performed with 2% FBS-PBS.

    Cell Cycle Assay:

    Article Title: Trans‐gnetin H isolated from the seeds of Paeonia species induces autophagy via inhibiting mTORC1 signalling through AMPK activation
    Article Snippet: The WB protocol was the same as our previous work.The WB protocol was the same as our previous work.. The primary antibodies Actin (20536‐1‐AP), LC3II (18725‐1‐AP), p62 (18420‐1‐AP) were purchased from Proteintech; TFEB (37785 S), pT389‐S6K (9234 S), p‐S6 (4858 S), S6K (9202 S), S6 (2217 S), p‐AMPKα (50081 S), AMPKα (5832 S), Raptor (2280 S) and TSC2 (D93F12) were purchased from Cell Signaling Technology; Flag (db7002) and HA (db2603) were purchased from Hangzhou Bio Technolog.. The secondary antibodies were obtained from Sigma.The secondary antibodies were obtained from Sigma.

    Article Title: A translational program that suppresses metabolism to shield the genome
    Article Snippet: Blots were stripped and reprobed when looking at total versus modified proteins unless otherwise noted.. Antibodies (all 1:1000): GLUT1 (Novus Biologicals, # NB110-39113), PAI-1 (Proteintech, #13801-1-AP), Puromycin (3RH11) (Kerafast, #EQ0001), β-ACTIN (C4) (SantaCruz Biotechnology, #sc-47778), EIF5A (abcam, #ab32443), EIF3D (Proteintech, # 10219-1-AP), EIF3K (Proteintech, # 10640-1-AP), acetyl-EIF5A (Lys47) (Boster Bio, #P01727), CYR61 (Proteintech, #26689-1-AP), SDC4 (R&D Systems, #P31431), SIRT1 (Proteintech, #13161-1-AP), SIRT2 (Proteintech, #19655-1-AP), NFKB2 (Proteintech, #10409-2-AP), 4E-BP (Cell Signaling, #9452 S), Phospho-4E-BP (Ser65) (Cell Signaling, #9451 S), TSC2 (D93F12) (Cell Signaling, #4308S), gamma H2AX (Novus, NB100-384), H2AX (Abcam, Ab11175), CRM-1 (Novus, NB100-79802), XPO-4 (Abcam, ab133237), eIF5A2 (Thermo, #PA5-30770), Phospho-RAPTOR (Ser792) (Cell Signaling, #89146S), RAPTOR (Cell Signaling, #2280), Phospho-Tuberin/TSC2 (Ser1387) (Cell Signaling, #5584S), NDRG1 (Abcam, #ab37897), and Anti-hypusine (mAbHpu24) antibody was kindly provided by Genentech.. The eIF5A1 antibodies and siRNAs used in this study were specific to eIF5A1 and not eIF5A2 as described by the manufacturer and validated in Supplementary Fig. .

    Article Title: Identification of mechanically regulated phosphorylation sites on tuberin (TSC2) that control mechanistic target of rapamycin (mTOR) signaling
    Article Snippet: Images were quantified using ImageJ software (U.S. National Institutes of Health, Bethesda, MD; http://rsb.info.nih.gov/nih-image/).Images were quantified using ImageJ software (U.S. National Institutes of Health, Bethesda, MD; http://rsb.info.nih.gov/nih-image/).. Antibodies - P-p70(389) (1A5) (#9206, used to probe Myc-tagged P-p70), P-p70(389) (#9234, used to probe endogenous P-p70), total p70 (49D7) (#2708), Anti-P-Akt substrate (RxRxxS*/T*) (23C8D2) for detection of PTSC2 (#10001), P-JNK(Thr183/Tyr185) (98F2) (#4671), total JNK (#9252), total mTOR (#2972), total Rictor (D16H9) (#9476), P-PKB (Ser473) (#4060), P-PKB (Thr308) (#9275S), total PKB (#9272), P-S6 (Ser240/4) (#5364), P-S6 (Ser235/6) (#2211), total S6 (#2217S), total TSC1 (D43E2) (#6935), total TSC2 (D93F12) (#4308) were purchased from Cell Signaling Technology (Danvers, MA, USA).. Anti-puromycin (12D10) (#MABE343) was purchased from EMD Millipore (Billerica, MA, USA).Anti-puromycin (12D10) (#MABE343) was purchased from EMD Millipore (Billerica, MA, USA).

    Article Title: Chronic signaling via the metabolic checkpoint kinase mTORC1 induces macrophage granuloma formation and marks sarcoidosis progression
    Article Snippet: HRP-conjugated secondary antibodies (Bethyl) 1:10000 were applied for 1 hour at room temperature in 4% low-fat milk and proteins were visualized using ECL substrate (Thermo Scientific).HRP-conjugated secondary antibodies (Bethyl) 1:10000 were applied for 1 hour at room temperature in 4% low-fat milk and proteins were visualized using ECL substrate (Thermo Scientific).. Primary antibodies were TSC2 (D93F12) 1:1000, p-Akt S473 (D9E) 1:1000, 4E-BP1 (53H11) 1:1000, p-S6 S240/244 (rabbit polyclonal) 1:3000, S6 (54D2) 1:1000, p-Akt T308 (D25E6) 1:1000, pan-Akt (40D4) 1:1000, β-tubulin (9F3) 1:1000 , p-RB Ser807/811 (D20B12) 1:1000, PDCD4 (D29C6) 1:1000, cleaved caspase 3 Asp175 (5A1E or rabbit polyclonal) 1:500, survivin (71G4B7) 1:1000 and p-S6K1 T389 (108D2) 1:1000, p-TSC2 S939 (rabbit polyclonal) 1:1000 obtained from Cell Signaling Technology.. Bcl-2 (N-19) 1:500, cyclin D1 (M-20) 1:500, p27Kip1 (C-19) 1:1000, cyclin A (C-19) 1:500, CDK4 (C-22) 1:1000, CDK6 (H-96) 1:500, and E2F1 (KH95) 1:500 were fom Santa Cruz. α-tubulin (CP06) 1:3000 was purchased from Calbiochem and GAPDH (rabbit polyclonal) 1:10000 from Trevigen.Bcl-2 (N-19) 1:500, cyclin D1 (M-20) 1:500, p27Kip1 (C-19) 1:1000, cyclin A (C-19) 1:500, CDK4 (C-22) 1:1000, CDK6 (H-96) 1:500, and E2F1 (KH95) 1:500 were fom Santa Cruz. α-tubulin (CP06) 1:3000 was purchased..

    Article Title: TSC1 promotes i NKT cell anergy and inhibits i NKT cell-mediated anti-tumor immunity
    Article Snippet: Fluorescence-conjugated anti-mouse TCRβ (H57-597), NK1.1 (PK136), CD44(IM7), B220(RA3-6B2), Gr-1(RB6-8C5), CD11c(N418), CD11b(M1/70), CD4 (GK1.5), CD8 (53-6.7), IFN-γ (XMG1.2), IL-4 (11B11), PD-1(RMP1-30), and ICOS(D10.G4.1) antibodies were purchased from BioLegend.Fluorescence-conjugated anti-mouse TCRβ (H57-597), NK1.1 (PK136), CD44(IM7), B220(RA3-6B2), Gr-1(RB6-8C5), CD11c(N418), CD11b(M1/70), CD4 (GK1.5), CD8 (53-6.7), IFN-γ (XMG1.2), IL-4 (11B11), PD-1(RMP1-30), and ICOS(D10.G4.1) antibodies were purchased from BioLegend.. Anti–phospho-p44/42 mitogen-activated protein kinase T202/Y204 (197G2), anti–phospho-S6 S240/244 (D68F8), anti–phospho-4E-BP1 T37/46 (236B4), anti–phospho-Akt S473 (193H12), TSC1 (D43E2), and TSC2 (D93F12) were purchased from Cell Signaling Technology, Inc. Anti–β-actin antibody was purchased from Sigma-Aldrich.. Cell surface staining was performed with 2% FBS-PBS.Cell surface staining was performed with 2% FBS-PBS.

    Article Title: TSC1 promotes i NKT cell anergy and inhibits i NKT cell-mediated anti-tumor immunity
    Article Snippet: CD11b(M1/70), CD4 (GK1.5), CD8 (53-6.7), IFN-γ (XMG1.2), IL-4 (11B11), PD-1(RMP1-30), and ICOS(D10.G4.1) antibodies were purchased from BioLegend. .. Anti–phospho-p44/42 mitogen-activated protein kinase T202/Y204 (197G2), anti–phospho-S6 S240/244 (D68F8), anti–phospho-4E-BP1 T37/46 (236B4), anti–phospho-Akt S473 (193H12), TSC1 (D43E2), and TSC2 (D93F12) were purchased from Cell Signaling Technology, Inc. Anti–β-actin antibody was purchased from Sigma-Aldrich. .. Cell surface staining was performed with 2% FBS-PBS.

    Gene Expression:

    Article Title: Trans‐gnetin H isolated from the seeds of Paeonia species induces autophagy via inhibiting mTORC1 signalling through AMPK activation
    Article Snippet: The WB protocol was the same as our previous work.The WB protocol was the same as our previous work.. The primary antibodies Actin (20536‐1‐AP), LC3II (18725‐1‐AP), p62 (18420‐1‐AP) were purchased from Proteintech; TFEB (37785 S), pT389‐S6K (9234 S), p‐S6 (4858 S), S6K (9202 S), S6 (2217 S), p‐AMPKα (50081 S), AMPKα (5832 S), Raptor (2280 S) and TSC2 (D93F12) were purchased from Cell Signaling Technology; Flag (db7002) and HA (db2603) were purchased from Hangzhou Bio Technolog.. The secondary antibodies were obtained from Sigma.The secondary antibodies were obtained from Sigma.

    Article Title: A translational program that suppresses metabolism to shield the genome
    Article Snippet: Blots were stripped and reprobed when looking at total versus modified proteins unless otherwise noted.. Antibodies (all 1:1000): GLUT1 (Novus Biologicals, # NB110-39113), PAI-1 (Proteintech, #13801-1-AP), Puromycin (3RH11) (Kerafast, #EQ0001), β-ACTIN (C4) (SantaCruz Biotechnology, #sc-47778), EIF5A (abcam, #ab32443), EIF3D (Proteintech, # 10219-1-AP), EIF3K (Proteintech, # 10640-1-AP), acetyl-EIF5A (Lys47) (Boster Bio, #P01727), CYR61 (Proteintech, #26689-1-AP), SDC4 (R&D Systems, #P31431), SIRT1 (Proteintech, #13161-1-AP), SIRT2 (Proteintech, #19655-1-AP), NFKB2 (Proteintech, #10409-2-AP), 4E-BP (Cell Signaling, #9452 S), Phospho-4E-BP (Ser65) (Cell Signaling, #9451 S), TSC2 (D93F12) (Cell Signaling, #4308S), gamma H2AX (Novus, NB100-384), H2AX (Abcam, Ab11175), CRM-1 (Novus, NB100-79802), XPO-4 (Abcam, ab133237), eIF5A2 (Thermo, #PA5-30770), Phospho-RAPTOR (Ser792) (Cell Signaling, #89146S), RAPTOR (Cell Signaling, #2280), Phospho-Tuberin/TSC2 (Ser1387) (Cell Signaling, #5584S), NDRG1 (Abcam, #ab37897), and Anti-hypusine (mAbHpu24) antibody was kindly provided by Genentech.. The eIF5A1 antibodies and siRNAs used in this study were specific to eIF5A1 and not eIF5A2 as described by the manufacturer and validated in Supplementary Fig. .

    Article Title: Identification of mechanically regulated phosphorylation sites on tuberin (TSC2) that control mechanistic target of rapamycin (mTOR) signaling
    Article Snippet: Images were quantified using ImageJ software (U.S. National Institutes of Health, Bethesda, MD; http://rsb.info.nih.gov/nih-image/).Images were quantified using ImageJ software (U.S. National Institutes of Health, Bethesda, MD; http://rsb.info.nih.gov/nih-image/).. Antibodies - P-p70(389) (1A5) (#9206, used to probe Myc-tagged P-p70), P-p70(389) (#9234, used to probe endogenous P-p70), total p70 (49D7) (#2708), Anti-P-Akt substrate (RxRxxS*/T*) (23C8D2) for detection of PTSC2 (#10001), P-JNK(Thr183/Tyr185) (98F2) (#4671), total JNK (#9252), total mTOR (#2972), total Rictor (D16H9) (#9476), P-PKB (Ser473) (#4060), P-PKB (Thr308) (#9275S), total PKB (#9272), P-S6 (Ser240/4) (#5364), P-S6 (Ser235/6) (#2211), total S6 (#2217S), total TSC1 (D43E2) (#6935), total TSC2 (D93F12) (#4308) were purchased from Cell Signaling Technology (Danvers, MA, USA).. Anti-puromycin (12D10) (#MABE343) was purchased from EMD Millipore (Billerica, MA, USA).Anti-puromycin (12D10) (#MABE343) was purchased from EMD Millipore (Billerica, MA, USA).

    Article Title: Chronic signaling via the metabolic checkpoint kinase mTORC1 induces macrophage granuloma formation and marks sarcoidosis progression
    Article Snippet: HRP-conjugated secondary antibodies (Bethyl) 1:10000 were applied for 1 hour at room temperature in 4% low-fat milk and proteins were visualized using ECL substrate (Thermo Scientific).HRP-conjugated secondary antibodies (Bethyl) 1:10000 were applied for 1 hour at room temperature in 4% low-fat milk and proteins were visualized using ECL substrate (Thermo Scientific).. Primary antibodies were TSC2 (D93F12) 1:1000, p-Akt S473 (D9E) 1:1000, 4E-BP1 (53H11) 1:1000, p-S6 S240/244 (rabbit polyclonal) 1:3000, S6 (54D2) 1:1000, p-Akt T308 (D25E6) 1:1000, pan-Akt (40D4) 1:1000, β-tubulin (9F3) 1:1000 , p-RB Ser807/811 (D20B12) 1:1000, PDCD4 (D29C6) 1:1000, cleaved caspase 3 Asp175 (5A1E or rabbit polyclonal) 1:500, survivin (71G4B7) 1:1000 and p-S6K1 T389 (108D2) 1:1000, p-TSC2 S939 (rabbit polyclonal) 1:1000 obtained from Cell Signaling Technology.. Bcl-2 (N-19) 1:500, cyclin D1 (M-20) 1:500, p27Kip1 (C-19) 1:1000, cyclin A (C-19) 1:500, CDK4 (C-22) 1:1000, CDK6 (H-96) 1:500, and E2F1 (KH95) 1:500 were fom Santa Cruz. α-tubulin (CP06) 1:3000 was purchased from Calbiochem and GAPDH (rabbit polyclonal) 1:10000 from Trevigen.Bcl-2 (N-19) 1:500, cyclin D1 (M-20) 1:500, p27Kip1 (C-19) 1:1000, cyclin A (C-19) 1:500, CDK4 (C-22) 1:1000, CDK6 (H-96) 1:500, and E2F1 (KH95) 1:500 were fom Santa Cruz. α-tubulin (CP06) 1:3000 was purchased..

    Article Title: TSC1 promotes i NKT cell anergy and inhibits i NKT cell-mediated anti-tumor immunity
    Article Snippet: Fluorescence-conjugated anti-mouse TCRβ (H57-597), NK1.1 (PK136), CD44(IM7), B220(RA3-6B2), Gr-1(RB6-8C5), CD11c(N418), CD11b(M1/70), CD4 (GK1.5), CD8 (53-6.7), IFN-γ (XMG1.2), IL-4 (11B11), PD-1(RMP1-30), and ICOS(D10.G4.1) antibodies were purchased from BioLegend.Fluorescence-conjugated anti-mouse TCRβ (H57-597), NK1.1 (PK136), CD44(IM7), B220(RA3-6B2), Gr-1(RB6-8C5), CD11c(N418), CD11b(M1/70), CD4 (GK1.5), CD8 (53-6.7), IFN-γ (XMG1.2), IL-4 (11B11), PD-1(RMP1-30), and ICOS(D10.G4.1) antibodies were purchased from BioLegend.. Anti–phospho-p44/42 mitogen-activated protein kinase T202/Y204 (197G2), anti–phospho-S6 S240/244 (D68F8), anti–phospho-4E-BP1 T37/46 (236B4), anti–phospho-Akt S473 (193H12), TSC1 (D43E2), and TSC2 (D93F12) were purchased from Cell Signaling Technology, Inc. Anti–β-actin antibody was purchased from Sigma-Aldrich.. Cell surface staining was performed with 2% FBS-PBS.Cell surface staining was performed with 2% FBS-PBS.

    Article Title: TSC1 promotes i NKT cell anergy and inhibits i NKT cell-mediated anti-tumor immunity
    Article Snippet: CD11b(M1/70), CD4 (GK1.5), CD8 (53-6.7), IFN-γ (XMG1.2), IL-4 (11B11), PD-1(RMP1-30), and ICOS(D10.G4.1) antibodies were purchased from BioLegend. .. Anti–phospho-p44/42 mitogen-activated protein kinase T202/Y204 (197G2), anti–phospho-S6 S240/244 (D68F8), anti–phospho-4E-BP1 T37/46 (236B4), anti–phospho-Akt S473 (193H12), TSC1 (D43E2), and TSC2 (D93F12) were purchased from Cell Signaling Technology, Inc. Anti–β-actin antibody was purchased from Sigma-Aldrich. .. Cell surface staining was performed with 2% FBS-PBS.

    Activity Assay:

    Article Title: Trans‐gnetin H isolated from the seeds of Paeonia species induces autophagy via inhibiting mTORC1 signalling through AMPK activation
    Article Snippet: The WB protocol was the same as our previous work.The WB protocol was the same as our previous work.. The primary antibodies Actin (20536‐1‐AP), LC3II (18725‐1‐AP), p62 (18420‐1‐AP) were purchased from Proteintech; TFEB (37785 S), pT389‐S6K (9234 S), p‐S6 (4858 S), S6K (9202 S), S6 (2217 S), p‐AMPKα (50081 S), AMPKα (5832 S), Raptor (2280 S) and TSC2 (D93F12) were purchased from Cell Signaling Technology; Flag (db7002) and HA (db2603) were purchased from Hangzhou Bio Technolog.. The secondary antibodies were obtained from Sigma.The secondary antibodies were obtained from Sigma.

    Article Title: A translational program that suppresses metabolism to shield the genome
    Article Snippet: Blots were stripped and reprobed when looking at total versus modified proteins unless otherwise noted.. Antibodies (all 1:1000): GLUT1 (Novus Biologicals, # NB110-39113), PAI-1 (Proteintech, #13801-1-AP), Puromycin (3RH11) (Kerafast, #EQ0001), β-ACTIN (C4) (SantaCruz Biotechnology, #sc-47778), EIF5A (abcam, #ab32443), EIF3D (Proteintech, # 10219-1-AP), EIF3K (Proteintech, # 10640-1-AP), acetyl-EIF5A (Lys47) (Boster Bio, #P01727), CYR61 (Proteintech, #26689-1-AP), SDC4 (R&D Systems, #P31431), SIRT1 (Proteintech, #13161-1-AP), SIRT2 (Proteintech, #19655-1-AP), NFKB2 (Proteintech, #10409-2-AP), 4E-BP (Cell Signaling, #9452 S), Phospho-4E-BP (Ser65) (Cell Signaling, #9451 S), TSC2 (D93F12) (Cell Signaling, #4308S), gamma H2AX (Novus, NB100-384), H2AX (Abcam, Ab11175), CRM-1 (Novus, NB100-79802), XPO-4 (Abcam, ab133237), eIF5A2 (Thermo, #PA5-30770), Phospho-RAPTOR (Ser792) (Cell Signaling, #89146S), RAPTOR (Cell Signaling, #2280), Phospho-Tuberin/TSC2 (Ser1387) (Cell Signaling, #5584S), NDRG1 (Abcam, #ab37897), and Anti-hypusine (mAbHpu24) antibody was kindly provided by Genentech.. The eIF5A1 antibodies and siRNAs used in this study were specific to eIF5A1 and not eIF5A2 as described by the manufacturer and validated in Supplementary Fig. .

    Article Title: Identification of mechanically regulated phosphorylation sites on tuberin (TSC2) that control mechanistic target of rapamycin (mTOR) signaling
    Article Snippet: Images were quantified using ImageJ software (U.S. National Institutes of Health, Bethesda, MD; http://rsb.info.nih.gov/nih-image/).Images were quantified using ImageJ software (U.S. National Institutes of Health, Bethesda, MD; http://rsb.info.nih.gov/nih-image/).. Antibodies - P-p70(389) (1A5) (#9206, used to probe Myc-tagged P-p70), P-p70(389) (#9234, used to probe endogenous P-p70), total p70 (49D7) (#2708), Anti-P-Akt substrate (RxRxxS*/T*) (23C8D2) for detection of PTSC2 (#10001), P-JNK(Thr183/Tyr185) (98F2) (#4671), total JNK (#9252), total mTOR (#2972), total Rictor (D16H9) (#9476), P-PKB (Ser473) (#4060), P-PKB (Thr308) (#9275S), total PKB (#9272), P-S6 (Ser240/4) (#5364), P-S6 (Ser235/6) (#2211), total S6 (#2217S), total TSC1 (D43E2) (#6935), total TSC2 (D93F12) (#4308) were purchased from Cell Signaling Technology (Danvers, MA, USA).. Anti-puromycin (12D10) (#MABE343) was purchased from EMD Millipore (Billerica, MA, USA).Anti-puromycin (12D10) (#MABE343) was purchased from EMD Millipore (Billerica, MA, USA).

    Article Title: Chronic signaling via the metabolic checkpoint kinase mTORC1 induces macrophage granuloma formation and marks sarcoidosis progression
    Article Snippet: HRP-conjugated secondary antibodies (Bethyl) 1:10000 were applied for 1 hour at room temperature in 4% low-fat milk and proteins were visualized using ECL substrate (Thermo Scientific).HRP-conjugated secondary antibodies (Bethyl) 1:10000 were applied for 1 hour at room temperature in 4% low-fat milk and proteins were visualized using ECL substrate (Thermo Scientific).. Primary antibodies were TSC2 (D93F12) 1:1000, p-Akt S473 (D9E) 1:1000, 4E-BP1 (53H11) 1:1000, p-S6 S240/244 (rabbit polyclonal) 1:3000, S6 (54D2) 1:1000, p-Akt T308 (D25E6) 1:1000, pan-Akt (40D4) 1:1000, β-tubulin (9F3) 1:1000 , p-RB Ser807/811 (D20B12) 1:1000, PDCD4 (D29C6) 1:1000, cleaved caspase 3 Asp175 (5A1E or rabbit polyclonal) 1:500, survivin (71G4B7) 1:1000 and p-S6K1 T389 (108D2) 1:1000, p-TSC2 S939 (rabbit polyclonal) 1:1000 obtained from Cell Signaling Technology.. Bcl-2 (N-19) 1:500, cyclin D1 (M-20) 1:500, p27Kip1 (C-19) 1:1000, cyclin A (C-19) 1:500, CDK4 (C-22) 1:1000, CDK6 (H-96) 1:500, and E2F1 (KH95) 1:500 were fom Santa Cruz. α-tubulin (CP06) 1:3000 was purchased from Calbiochem and GAPDH (rabbit polyclonal) 1:10000 from Trevigen.Bcl-2 (N-19) 1:500, cyclin D1 (M-20) 1:500, p27Kip1 (C-19) 1:1000, cyclin A (C-19) 1:500, CDK4 (C-22) 1:1000, CDK6 (H-96) 1:500, and E2F1 (KH95) 1:500 were fom Santa Cruz. α-tubulin (CP06) 1:3000 was purchased..

    Article Title: TSC1 promotes i NKT cell anergy and inhibits i NKT cell-mediated anti-tumor immunity
    Article Snippet: Fluorescence-conjugated anti-mouse TCRβ (H57-597), NK1.1 (PK136), CD44(IM7), B220(RA3-6B2), Gr-1(RB6-8C5), CD11c(N418), CD11b(M1/70), CD4 (GK1.5), CD8 (53-6.7), IFN-γ (XMG1.2), IL-4 (11B11), PD-1(RMP1-30), and ICOS(D10.G4.1) antibodies were purchased from BioLegend.Fluorescence-conjugated anti-mouse TCRβ (H57-597), NK1.1 (PK136), CD44(IM7), B220(RA3-6B2), Gr-1(RB6-8C5), CD11c(N418), CD11b(M1/70), CD4 (GK1.5), CD8 (53-6.7), IFN-γ (XMG1.2), IL-4 (11B11), PD-1(RMP1-30), and ICOS(D10.G4.1) antibodies were purchased from BioLegend.. Anti–phospho-p44/42 mitogen-activated protein kinase T202/Y204 (197G2), anti–phospho-S6 S240/244 (D68F8), anti–phospho-4E-BP1 T37/46 (236B4), anti–phospho-Akt S473 (193H12), TSC1 (D43E2), and TSC2 (D93F12) were purchased from Cell Signaling Technology, Inc. Anti–β-actin antibody was purchased from Sigma-Aldrich.. Cell surface staining was performed with 2% FBS-PBS.Cell surface staining was performed with 2% FBS-PBS.

    Article Title: TSC1 promotes i NKT cell anergy and inhibits i NKT cell-mediated anti-tumor immunity
    Article Snippet: CD11b(M1/70), CD4 (GK1.5), CD8 (53-6.7), IFN-γ (XMG1.2), IL-4 (11B11), PD-1(RMP1-30), and ICOS(D10.G4.1) antibodies were purchased from BioLegend. .. Anti–phospho-p44/42 mitogen-activated protein kinase T202/Y204 (197G2), anti–phospho-S6 S240/244 (D68F8), anti–phospho-4E-BP1 T37/46 (236B4), anti–phospho-Akt S473 (193H12), TSC1 (D43E2), and TSC2 (D93F12) were purchased from Cell Signaling Technology, Inc. Anti–β-actin antibody was purchased from Sigma-Aldrich. .. Cell surface staining was performed with 2% FBS-PBS.

    In Situ:

    Article Title: Trans‐gnetin H isolated from the seeds of Paeonia species induces autophagy via inhibiting mTORC1 signalling through AMPK activation
    Article Snippet: The WB protocol was the same as our previous work.The WB protocol was the same as our previous work.. The primary antibodies Actin (20536‐1‐AP), LC3II (18725‐1‐AP), p62 (18420‐1‐AP) were purchased from Proteintech; TFEB (37785 S), pT389‐S6K (9234 S), p‐S6 (4858 S), S6K (9202 S), S6 (2217 S), p‐AMPKα (50081 S), AMPKα (5832 S), Raptor (2280 S) and TSC2 (D93F12) were purchased from Cell Signaling Technology; Flag (db7002) and HA (db2603) were purchased from Hangzhou Bio Technolog.. The secondary antibodies were obtained from Sigma.The secondary antibodies were obtained from Sigma.

    Article Title: A translational program that suppresses metabolism to shield the genome
    Article Snippet: Blots were stripped and reprobed when looking at total versus modified proteins unless otherwise noted.. Antibodies (all 1:1000): GLUT1 (Novus Biologicals, # NB110-39113), PAI-1 (Proteintech, #13801-1-AP), Puromycin (3RH11) (Kerafast, #EQ0001), β-ACTIN (C4) (SantaCruz Biotechnology, #sc-47778), EIF5A (abcam, #ab32443), EIF3D (Proteintech, # 10219-1-AP), EIF3K (Proteintech, # 10640-1-AP), acetyl-EIF5A (Lys47) (Boster Bio, #P01727), CYR61 (Proteintech, #26689-1-AP), SDC4 (R&D Systems, #P31431), SIRT1 (Proteintech, #13161-1-AP), SIRT2 (Proteintech, #19655-1-AP), NFKB2 (Proteintech, #10409-2-AP), 4E-BP (Cell Signaling, #9452 S), Phospho-4E-BP (Ser65) (Cell Signaling, #9451 S), TSC2 (D93F12) (Cell Signaling, #4308S), gamma H2AX (Novus, NB100-384), H2AX (Abcam, Ab11175), CRM-1 (Novus, NB100-79802), XPO-4 (Abcam, ab133237), eIF5A2 (Thermo, #PA5-30770), Phospho-RAPTOR (Ser792) (Cell Signaling, #89146S), RAPTOR (Cell Signaling, #2280), Phospho-Tuberin/TSC2 (Ser1387) (Cell Signaling, #5584S), NDRG1 (Abcam, #ab37897), and Anti-hypusine (mAbHpu24) antibody was kindly provided by Genentech.. The eIF5A1 antibodies and siRNAs used in this study were specific to eIF5A1 and not eIF5A2 as described by the manufacturer and validated in Supplementary Fig. .

    Article Title: Identification of mechanically regulated phosphorylation sites on tuberin (TSC2) that control mechanistic target of rapamycin (mTOR) signaling
    Article Snippet: Images were quantified using ImageJ software (U.S. National Institutes of Health, Bethesda, MD; http://rsb.info.nih.gov/nih-image/).Images were quantified using ImageJ software (U.S. National Institutes of Health, Bethesda, MD; http://rsb.info.nih.gov/nih-image/).. Antibodies - P-p70(389) (1A5) (#9206, used to probe Myc-tagged P-p70), P-p70(389) (#9234, used to probe endogenous P-p70), total p70 (49D7) (#2708), Anti-P-Akt substrate (RxRxxS*/T*) (23C8D2) for detection of PTSC2 (#10001), P-JNK(Thr183/Tyr185) (98F2) (#4671), total JNK (#9252), total mTOR (#2972), total Rictor (D16H9) (#9476), P-PKB (Ser473) (#4060), P-PKB (Thr308) (#9275S), total PKB (#9272), P-S6 (Ser240/4) (#5364), P-S6 (Ser235/6) (#2211), total S6 (#2217S), total TSC1 (D43E2) (#6935), total TSC2 (D93F12) (#4308) were purchased from Cell Signaling Technology (Danvers, MA, USA).. Anti-puromycin (12D10) (#MABE343) was purchased from EMD Millipore (Billerica, MA, USA).Anti-puromycin (12D10) (#MABE343) was purchased from EMD Millipore (Billerica, MA, USA).

    Article Title: Chronic signaling via the metabolic checkpoint kinase mTORC1 induces macrophage granuloma formation and marks sarcoidosis progression
    Article Snippet: HRP-conjugated secondary antibodies (Bethyl) 1:10000 were applied for 1 hour at room temperature in 4% low-fat milk and proteins were visualized using ECL substrate (Thermo Scientific).HRP-conjugated secondary antibodies (Bethyl) 1:10000 were applied for 1 hour at room temperature in 4% low-fat milk and proteins were visualized using ECL substrate (Thermo Scientific).. Primary antibodies were TSC2 (D93F12) 1:1000, p-Akt S473 (D9E) 1:1000, 4E-BP1 (53H11) 1:1000, p-S6 S240/244 (rabbit polyclonal) 1:3000, S6 (54D2) 1:1000, p-Akt T308 (D25E6) 1:1000, pan-Akt (40D4) 1:1000, β-tubulin (9F3) 1:1000 , p-RB Ser807/811 (D20B12) 1:1000, PDCD4 (D29C6) 1:1000, cleaved caspase 3 Asp175 (5A1E or rabbit polyclonal) 1:500, survivin (71G4B7) 1:1000 and p-S6K1 T389 (108D2) 1:1000, p-TSC2 S939 (rabbit polyclonal) 1:1000 obtained from Cell Signaling Technology.. Bcl-2 (N-19) 1:500, cyclin D1 (M-20) 1:500, p27Kip1 (C-19) 1:1000, cyclin A (C-19) 1:500, CDK4 (C-22) 1:1000, CDK6 (H-96) 1:500, and E2F1 (KH95) 1:500 were fom Santa Cruz. α-tubulin (CP06) 1:3000 was purchased from Calbiochem and GAPDH (rabbit polyclonal) 1:10000 from Trevigen.Bcl-2 (N-19) 1:500, cyclin D1 (M-20) 1:500, p27Kip1 (C-19) 1:1000, cyclin A (C-19) 1:500, CDK4 (C-22) 1:1000, CDK6 (H-96) 1:500, and E2F1 (KH95) 1:500 were fom Santa Cruz. α-tubulin (CP06) 1:3000 was purchased..

    Article Title: TSC1 promotes i NKT cell anergy and inhibits i NKT cell-mediated anti-tumor immunity
    Article Snippet: Fluorescence-conjugated anti-mouse TCRβ (H57-597), NK1.1 (PK136), CD44(IM7), B220(RA3-6B2), Gr-1(RB6-8C5), CD11c(N418), CD11b(M1/70), CD4 (GK1.5), CD8 (53-6.7), IFN-γ (XMG1.2), IL-4 (11B11), PD-1(RMP1-30), and ICOS(D10.G4.1) antibodies were purchased from BioLegend.Fluorescence-conjugated anti-mouse TCRβ (H57-597), NK1.1 (PK136), CD44(IM7), B220(RA3-6B2), Gr-1(RB6-8C5), CD11c(N418), CD11b(M1/70), CD4 (GK1.5), CD8 (53-6.7), IFN-γ (XMG1.2), IL-4 (11B11), PD-1(RMP1-30), and ICOS(D10.G4.1) antibodies were purchased from BioLegend.. Anti–phospho-p44/42 mitogen-activated protein kinase T202/Y204 (197G2), anti–phospho-S6 S240/244 (D68F8), anti–phospho-4E-BP1 T37/46 (236B4), anti–phospho-Akt S473 (193H12), TSC1 (D43E2), and TSC2 (D93F12) were purchased from Cell Signaling Technology, Inc. Anti–β-actin antibody was purchased from Sigma-Aldrich.. Cell surface staining was performed with 2% FBS-PBS.Cell surface staining was performed with 2% FBS-PBS.

    Article Title: TSC1 promotes i NKT cell anergy and inhibits i NKT cell-mediated anti-tumor immunity
    Article Snippet: CD11b(M1/70), CD4 (GK1.5), CD8 (53-6.7), IFN-γ (XMG1.2), IL-4 (11B11), PD-1(RMP1-30), and ICOS(D10.G4.1) antibodies were purchased from BioLegend. .. Anti–phospho-p44/42 mitogen-activated protein kinase T202/Y204 (197G2), anti–phospho-S6 S240/244 (D68F8), anti–phospho-4E-BP1 T37/46 (236B4), anti–phospho-Akt S473 (193H12), TSC1 (D43E2), and TSC2 (D93F12) were purchased from Cell Signaling Technology, Inc. Anti–β-actin antibody was purchased from Sigma-Aldrich. .. Cell surface staining was performed with 2% FBS-PBS.

    Microarray:

    Article Title: Trans‐gnetin H isolated from the seeds of Paeonia species induces autophagy via inhibiting mTORC1 signalling through AMPK activation
    Article Snippet: The WB protocol was the same as our previous work.The WB protocol was the same as our previous work.. The primary antibodies Actin (20536‐1‐AP), LC3II (18725‐1‐AP), p62 (18420‐1‐AP) were purchased from Proteintech; TFEB (37785 S), pT389‐S6K (9234 S), p‐S6 (4858 S), S6K (9202 S), S6 (2217 S), p‐AMPKα (50081 S), AMPKα (5832 S), Raptor (2280 S) and TSC2 (D93F12) were purchased from Cell Signaling Technology; Flag (db7002) and HA (db2603) were purchased from Hangzhou Bio Technolog.. The secondary antibodies were obtained from Sigma.The secondary antibodies were obtained from Sigma.

    Article Title: A translational program that suppresses metabolism to shield the genome
    Article Snippet: Blots were stripped and reprobed when looking at total versus modified proteins unless otherwise noted.. Antibodies (all 1:1000): GLUT1 (Novus Biologicals, # NB110-39113), PAI-1 (Proteintech, #13801-1-AP), Puromycin (3RH11) (Kerafast, #EQ0001), β-ACTIN (C4) (SantaCruz Biotechnology, #sc-47778), EIF5A (abcam, #ab32443), EIF3D (Proteintech, # 10219-1-AP), EIF3K (Proteintech, # 10640-1-AP), acetyl-EIF5A (Lys47) (Boster Bio, #P01727), CYR61 (Proteintech, #26689-1-AP), SDC4 (R&D Systems, #P31431), SIRT1 (Proteintech, #13161-1-AP), SIRT2 (Proteintech, #19655-1-AP), NFKB2 (Proteintech, #10409-2-AP), 4E-BP (Cell Signaling, #9452 S), Phospho-4E-BP (Ser65) (Cell Signaling, #9451 S), TSC2 (D93F12) (Cell Signaling, #4308S), gamma H2AX (Novus, NB100-384), H2AX (Abcam, Ab11175), CRM-1 (Novus, NB100-79802), XPO-4 (Abcam, ab133237), eIF5A2 (Thermo, #PA5-30770), Phospho-RAPTOR (Ser792) (Cell Signaling, #89146S), RAPTOR (Cell Signaling, #2280), Phospho-Tuberin/TSC2 (Ser1387) (Cell Signaling, #5584S), NDRG1 (Abcam, #ab37897), and Anti-hypusine (mAbHpu24) antibody was kindly provided by Genentech.. The eIF5A1 antibodies and siRNAs used in this study were specific to eIF5A1 and not eIF5A2 as described by the manufacturer and validated in Supplementary Fig. .

    Article Title: Identification of mechanically regulated phosphorylation sites on tuberin (TSC2) that control mechanistic target of rapamycin (mTOR) signaling
    Article Snippet: Images were quantified using ImageJ software (U.S. National Institutes of Health, Bethesda, MD; http://rsb.info.nih.gov/nih-image/).Images were quantified using ImageJ software (U.S. National Institutes of Health, Bethesda, MD; http://rsb.info.nih.gov/nih-image/).. Antibodies - P-p70(389) (1A5) (#9206, used to probe Myc-tagged P-p70), P-p70(389) (#9234, used to probe endogenous P-p70), total p70 (49D7) (#2708), Anti-P-Akt substrate (RxRxxS*/T*) (23C8D2) for detection of PTSC2 (#10001), P-JNK(Thr183/Tyr185) (98F2) (#4671), total JNK (#9252), total mTOR (#2972), total Rictor (D16H9) (#9476), P-PKB (Ser473) (#4060), P-PKB (Thr308) (#9275S), total PKB (#9272), P-S6 (Ser240/4) (#5364), P-S6 (Ser235/6) (#2211), total S6 (#2217S), total TSC1 (D43E2) (#6935), total TSC2 (D93F12) (#4308) were purchased from Cell Signaling Technology (Danvers, MA, USA).. Anti-puromycin (12D10) (#MABE343) was purchased from EMD Millipore (Billerica, MA, USA).Anti-puromycin (12D10) (#MABE343) was purchased from EMD Millipore (Billerica, MA, USA).

    Article Title: Chronic signaling via the metabolic checkpoint kinase mTORC1 induces macrophage granuloma formation and marks sarcoidosis progression
    Article Snippet: HRP-conjugated secondary antibodies (Bethyl) 1:10000 were applied for 1 hour at room temperature in 4% low-fat milk and proteins were visualized using ECL substrate (Thermo Scientific).HRP-conjugated secondary antibodies (Bethyl) 1:10000 were applied for 1 hour at room temperature in 4% low-fat milk and proteins were visualized using ECL substrate (Thermo Scientific).. Primary antibodies were TSC2 (D93F12) 1:1000, p-Akt S473 (D9E) 1:1000, 4E-BP1 (53H11) 1:1000, p-S6 S240/244 (rabbit polyclonal) 1:3000, S6 (54D2) 1:1000, p-Akt T308 (D25E6) 1:1000, pan-Akt (40D4) 1:1000, β-tubulin (9F3) 1:1000 , p-RB Ser807/811 (D20B12) 1:1000, PDCD4 (D29C6) 1:1000, cleaved caspase 3 Asp175 (5A1E or rabbit polyclonal) 1:500, survivin (71G4B7) 1:1000 and p-S6K1 T389 (108D2) 1:1000, p-TSC2 S939 (rabbit polyclonal) 1:1000 obtained from Cell Signaling Technology.. Bcl-2 (N-19) 1:500, cyclin D1 (M-20) 1:500, p27Kip1 (C-19) 1:1000, cyclin A (C-19) 1:500, CDK4 (C-22) 1:1000, CDK6 (H-96) 1:500, and E2F1 (KH95) 1:500 were fom Santa Cruz. α-tubulin (CP06) 1:3000 was purchased from Calbiochem and GAPDH (rabbit polyclonal) 1:10000 from Trevigen.Bcl-2 (N-19) 1:500, cyclin D1 (M-20) 1:500, p27Kip1 (C-19) 1:1000, cyclin A (C-19) 1:500, CDK4 (C-22) 1:1000, CDK6 (H-96) 1:500, and E2F1 (KH95) 1:500 were fom Santa Cruz. α-tubulin (CP06) 1:3000 was purchased..

    Article Title: TSC1 promotes i NKT cell anergy and inhibits i NKT cell-mediated anti-tumor immunity
    Article Snippet: Fluorescence-conjugated anti-mouse TCRβ (H57-597), NK1.1 (PK136), CD44(IM7), B220(RA3-6B2), Gr-1(RB6-8C5), CD11c(N418), CD11b(M1/70), CD4 (GK1.5), CD8 (53-6.7), IFN-γ (XMG1.2), IL-4 (11B11), PD-1(RMP1-30), and ICOS(D10.G4.1) antibodies were purchased from BioLegend.Fluorescence-conjugated anti-mouse TCRβ (H57-597), NK1.1 (PK136), CD44(IM7), B220(RA3-6B2), Gr-1(RB6-8C5), CD11c(N418), CD11b(M1/70), CD4 (GK1.5), CD8 (53-6.7), IFN-γ (XMG1.2), IL-4 (11B11), PD-1(RMP1-30), and ICOS(D10.G4.1) antibodies were purchased from BioLegend.. Anti–phospho-p44/42 mitogen-activated protein kinase T202/Y204 (197G2), anti–phospho-S6 S240/244 (D68F8), anti–phospho-4E-BP1 T37/46 (236B4), anti–phospho-Akt S473 (193H12), TSC1 (D43E2), and TSC2 (D93F12) were purchased from Cell Signaling Technology, Inc. Anti–β-actin antibody was purchased from Sigma-Aldrich.. Cell surface staining was performed with 2% FBS-PBS.Cell surface staining was performed with 2% FBS-PBS.

    Article Title: TSC1 promotes i NKT cell anergy and inhibits i NKT cell-mediated anti-tumor immunity
    Article Snippet: CD11b(M1/70), CD4 (GK1.5), CD8 (53-6.7), IFN-γ (XMG1.2), IL-4 (11B11), PD-1(RMP1-30), and ICOS(D10.G4.1) antibodies were purchased from BioLegend. .. Anti–phospho-p44/42 mitogen-activated protein kinase T202/Y204 (197G2), anti–phospho-S6 S240/244 (D68F8), anti–phospho-4E-BP1 T37/46 (236B4), anti–phospho-Akt S473 (193H12), TSC1 (D43E2), and TSC2 (D93F12) were purchased from Cell Signaling Technology, Inc. Anti–β-actin antibody was purchased from Sigma-Aldrich. .. Cell surface staining was performed with 2% FBS-PBS.

    Purification:

    Article Title: Trans‐gnetin H isolated from the seeds of Paeonia species induces autophagy via inhibiting mTORC1 signalling through AMPK activation
    Article Snippet: The WB protocol was the same as our previous work.The WB protocol was the same as our previous work.. The primary antibodies Actin (20536‐1‐AP), LC3II (18725‐1‐AP), p62 (18420‐1‐AP) were purchased from Proteintech; TFEB (37785 S), pT389‐S6K (9234 S), p‐S6 (4858 S), S6K (9202 S), S6 (2217 S), p‐AMPKα (50081 S), AMPKα (5832 S), Raptor (2280 S) and TSC2 (D93F12) were purchased from Cell Signaling Technology; Flag (db7002) and HA (db2603) were purchased from Hangzhou Bio Technolog.. The secondary antibodies were obtained from Sigma.The secondary antibodies were obtained from Sigma.

    Article Title: A translational program that suppresses metabolism to shield the genome
    Article Snippet: Blots were stripped and reprobed when looking at total versus modified proteins unless otherwise noted.. Antibodies (all 1:1000): GLUT1 (Novus Biologicals, # NB110-39113), PAI-1 (Proteintech, #13801-1-AP), Puromycin (3RH11) (Kerafast, #EQ0001), β-ACTIN (C4) (SantaCruz Biotechnology, #sc-47778), EIF5A (abcam, #ab32443), EIF3D (Proteintech, # 10219-1-AP), EIF3K (Proteintech, # 10640-1-AP), acetyl-EIF5A (Lys47) (Boster Bio, #P01727), CYR61 (Proteintech, #26689-1-AP), SDC4 (R&D Systems, #P31431), SIRT1 (Proteintech, #13161-1-AP), SIRT2 (Proteintech, #19655-1-AP), NFKB2 (Proteintech, #10409-2-AP), 4E-BP (Cell Signaling, #9452 S), Phospho-4E-BP (Ser65) (Cell Signaling, #9451 S), TSC2 (D93F12) (Cell Signaling, #4308S), gamma H2AX (Novus, NB100-384), H2AX (Abcam, Ab11175), CRM-1 (Novus, NB100-79802), XPO-4 (Abcam, ab133237), eIF5A2 (Thermo, #PA5-30770), Phospho-RAPTOR (Ser792) (Cell Signaling, #89146S), RAPTOR (Cell Signaling, #2280), Phospho-Tuberin/TSC2 (Ser1387) (Cell Signaling, #5584S), NDRG1 (Abcam, #ab37897), and Anti-hypusine (mAbHpu24) antibody was kindly provided by Genentech.. The eIF5A1 antibodies and siRNAs used in this study were specific to eIF5A1 and not eIF5A2 as described by the manufacturer and validated in Supplementary Fig. .

    Article Title: Identification of mechanically regulated phosphorylation sites on tuberin (TSC2) that control mechanistic target of rapamycin (mTOR) signaling
    Article Snippet: Images were quantified using ImageJ software (U.S. National Institutes of Health, Bethesda, MD; http://rsb.info.nih.gov/nih-image/).Images were quantified using ImageJ software (U.S. National Institutes of Health, Bethesda, MD; http://rsb.info.nih.gov/nih-image/).. Antibodies - P-p70(389) (1A5) (#9206, used to probe Myc-tagged P-p70), P-p70(389) (#9234, used to probe endogenous P-p70), total p70 (49D7) (#2708), Anti-P-Akt substrate (RxRxxS*/T*) (23C8D2) for detection of PTSC2 (#10001), P-JNK(Thr183/Tyr185) (98F2) (#4671), total JNK (#9252), total mTOR (#2972), total Rictor (D16H9) (#9476), P-PKB (Ser473) (#4060), P-PKB (Thr308) (#9275S), total PKB (#9272), P-S6 (Ser240/4) (#5364), P-S6 (Ser235/6) (#2211), total S6 (#2217S), total TSC1 (D43E2) (#6935), total TSC2 (D93F12) (#4308) were purchased from Cell Signaling Technology (Danvers, MA, USA).. Anti-puromycin (12D10) (#MABE343) was purchased from EMD Millipore (Billerica, MA, USA).Anti-puromycin (12D10) (#MABE343) was purchased from EMD Millipore (Billerica, MA, USA).

    Article Title: Chronic signaling via the metabolic checkpoint kinase mTORC1 induces macrophage granuloma formation and marks sarcoidosis progression
    Article Snippet: HRP-conjugated secondary antibodies (Bethyl) 1:10000 were applied for 1 hour at room temperature in 4% low-fat milk and proteins were visualized using ECL substrate (Thermo Scientific).HRP-conjugated secondary antibodies (Bethyl) 1:10000 were applied for 1 hour at room temperature in 4% low-fat milk and proteins were visualized using ECL substrate (Thermo Scientific).. Primary antibodies were TSC2 (D93F12) 1:1000, p-Akt S473 (D9E) 1:1000, 4E-BP1 (53H11) 1:1000, p-S6 S240/244 (rabbit polyclonal) 1:3000, S6 (54D2) 1:1000, p-Akt T308 (D25E6) 1:1000, pan-Akt (40D4) 1:1000, β-tubulin (9F3) 1:1000 , p-RB Ser807/811 (D20B12) 1:1000, PDCD4 (D29C6) 1:1000, cleaved caspase 3 Asp175 (5A1E or rabbit polyclonal) 1:500, survivin (71G4B7) 1:1000 and p-S6K1 T389 (108D2) 1:1000, p-TSC2 S939 (rabbit polyclonal) 1:1000 obtained from Cell Signaling Technology.. Bcl-2 (N-19) 1:500, cyclin D1 (M-20) 1:500, p27Kip1 (C-19) 1:1000, cyclin A (C-19) 1:500, CDK4 (C-22) 1:1000, CDK6 (H-96) 1:500, and E2F1 (KH95) 1:500 were fom Santa Cruz. α-tubulin (CP06) 1:3000 was purchased from Calbiochem and GAPDH (rabbit polyclonal) 1:10000 from Trevigen.Bcl-2 (N-19) 1:500, cyclin D1 (M-20) 1:500, p27Kip1 (C-19) 1:1000, cyclin A (C-19) 1:500, CDK4 (C-22) 1:1000, CDK6 (H-96) 1:500, and E2F1 (KH95) 1:500 were fom Santa Cruz. α-tubulin (CP06) 1:3000 was purchased..

    Article Title: TSC1 promotes i NKT cell anergy and inhibits i NKT cell-mediated anti-tumor immunity
    Article Snippet: Fluorescence-conjugated anti-mouse TCRβ (H57-597), NK1.1 (PK136), CD44(IM7), B220(RA3-6B2), Gr-1(RB6-8C5), CD11c(N418), CD11b(M1/70), CD4 (GK1.5), CD8 (53-6.7), IFN-γ (XMG1.2), IL-4 (11B11), PD-1(RMP1-30), and ICOS(D10.G4.1) antibodies were purchased from BioLegend.Fluorescence-conjugated anti-mouse TCRβ (H57-597), NK1.1 (PK136), CD44(IM7), B220(RA3-6B2), Gr-1(RB6-8C5), CD11c(N418), CD11b(M1/70), CD4 (GK1.5), CD8 (53-6.7), IFN-γ (XMG1.2), IL-4 (11B11), PD-1(RMP1-30), and ICOS(D10.G4.1) antibodies were purchased from BioLegend.. Anti–phospho-p44/42 mitogen-activated protein kinase T202/Y204 (197G2), anti–phospho-S6 S240/244 (D68F8), anti–phospho-4E-BP1 T37/46 (236B4), anti–phospho-Akt S473 (193H12), TSC1 (D43E2), and TSC2 (D93F12) were purchased from Cell Signaling Technology, Inc. Anti–β-actin antibody was purchased from Sigma-Aldrich.. Cell surface staining was performed with 2% FBS-PBS.Cell surface staining was performed with 2% FBS-PBS.

    Article Title: TSC1 promotes i NKT cell anergy and inhibits i NKT cell-mediated anti-tumor immunity
    Article Snippet: CD11b(M1/70), CD4 (GK1.5), CD8 (53-6.7), IFN-γ (XMG1.2), IL-4 (11B11), PD-1(RMP1-30), and ICOS(D10.G4.1) antibodies were purchased from BioLegend. .. Anti–phospho-p44/42 mitogen-activated protein kinase T202/Y204 (197G2), anti–phospho-S6 S240/244 (D68F8), anti–phospho-4E-BP1 T37/46 (236B4), anti–phospho-Akt S473 (193H12), TSC1 (D43E2), and TSC2 (D93F12) were purchased from Cell Signaling Technology, Inc. Anti–β-actin antibody was purchased from Sigma-Aldrich. .. Cell surface staining was performed with 2% FBS-PBS.

    Isolation:

    Article Title: Trans‐gnetin H isolated from the seeds of Paeonia species induces autophagy via inhibiting mTORC1 signalling through AMPK activation
    Article Snippet: The WB protocol was the same as our previous work.The WB protocol was the same as our previous work.. The primary antibodies Actin (20536‐1‐AP), LC3II (18725‐1‐AP), p62 (18420‐1‐AP) were purchased from Proteintech; TFEB (37785 S), pT389‐S6K (9234 S), p‐S6 (4858 S), S6K (9202 S), S6 (2217 S), p‐AMPKα (50081 S), AMPKα (5832 S), Raptor (2280 S) and TSC2 (D93F12) were purchased from Cell Signaling Technology; Flag (db7002) and HA (db2603) were purchased from Hangzhou Bio Technolog.. The secondary antibodies were obtained from Sigma.The secondary antibodies were obtained from Sigma.

    Article Title: A translational program that suppresses metabolism to shield the genome
    Article Snippet: Blots were stripped and reprobed when looking at total versus modified proteins unless otherwise noted.. Antibodies (all 1:1000): GLUT1 (Novus Biologicals, # NB110-39113), PAI-1 (Proteintech, #13801-1-AP), Puromycin (3RH11) (Kerafast, #EQ0001), β-ACTIN (C4) (SantaCruz Biotechnology, #sc-47778), EIF5A (abcam, #ab32443), EIF3D (Proteintech, # 10219-1-AP), EIF3K (Proteintech, # 10640-1-AP), acetyl-EIF5A (Lys47) (Boster Bio, #P01727), CYR61 (Proteintech, #26689-1-AP), SDC4 (R&D Systems, #P31431), SIRT1 (Proteintech, #13161-1-AP), SIRT2 (Proteintech, #19655-1-AP), NFKB2 (Proteintech, #10409-2-AP), 4E-BP (Cell Signaling, #9452 S), Phospho-4E-BP (Ser65) (Cell Signaling, #9451 S), TSC2 (D93F12) (Cell Signaling, #4308S), gamma H2AX (Novus, NB100-384), H2AX (Abcam, Ab11175), CRM-1 (Novus, NB100-79802), XPO-4 (Abcam, ab133237), eIF5A2 (Thermo, #PA5-30770), Phospho-RAPTOR (Ser792) (Cell Signaling, #89146S), RAPTOR (Cell Signaling, #2280), Phospho-Tuberin/TSC2 (Ser1387) (Cell Signaling, #5584S), NDRG1 (Abcam, #ab37897), and Anti-hypusine (mAbHpu24) antibody was kindly provided by Genentech.. The eIF5A1 antibodies and siRNAs used in this study were specific to eIF5A1 and not eIF5A2 as described by the manufacturer and validated in Supplementary Fig. .

    Article Title: Identification of mechanically regulated phosphorylation sites on tuberin (TSC2) that control mechanistic target of rapamycin (mTOR) signaling
    Article Snippet: Images were quantified using ImageJ software (U.S. National Institutes of Health, Bethesda, MD; http://rsb.info.nih.gov/nih-image/).Images were quantified using ImageJ software (U.S. National Institutes of Health, Bethesda, MD; http://rsb.info.nih.gov/nih-image/).. Antibodies - P-p70(389) (1A5) (#9206, used to probe Myc-tagged P-p70), P-p70(389) (#9234, used to probe endogenous P-p70), total p70 (49D7) (#2708), Anti-P-Akt substrate (RxRxxS*/T*) (23C8D2) for detection of PTSC2 (#10001), P-JNK(Thr183/Tyr185) (98F2) (#4671), total JNK (#9252), total mTOR (#2972), total Rictor (D16H9) (#9476), P-PKB (Ser473) (#4060), P-PKB (Thr308) (#9275S), total PKB (#9272), P-S6 (Ser240/4) (#5364), P-S6 (Ser235/6) (#2211), total S6 (#2217S), total TSC1 (D43E2) (#6935), total TSC2 (D93F12) (#4308) were purchased from Cell Signaling Technology (Danvers, MA, USA).. Anti-puromycin (12D10) (#MABE343) was purchased from EMD Millipore (Billerica, MA, USA).Anti-puromycin (12D10) (#MABE343) was purchased from EMD Millipore (Billerica, MA, USA).

    Article Title: Chronic signaling via the metabolic checkpoint kinase mTORC1 induces macrophage granuloma formation and marks sarcoidosis progression
    Article Snippet: HRP-conjugated secondary antibodies (Bethyl) 1:10000 were applied for 1 hour at room temperature in 4% low-fat milk and proteins were visualized using ECL substrate (Thermo Scientific).HRP-conjugated secondary antibodies (Bethyl) 1:10000 were applied for 1 hour at room temperature in 4% low-fat milk and proteins were visualized using ECL substrate (Thermo Scientific).. Primary antibodies were TSC2 (D93F12) 1:1000, p-Akt S473 (D9E) 1:1000, 4E-BP1 (53H11) 1:1000, p-S6 S240/244 (rabbit polyclonal) 1:3000, S6 (54D2) 1:1000, p-Akt T308 (D25E6) 1:1000, pan-Akt (40D4) 1:1000, β-tubulin (9F3) 1:1000 , p-RB Ser807/811 (D20B12) 1:1000, PDCD4 (D29C6) 1:1000, cleaved caspase 3 Asp175 (5A1E or rabbit polyclonal) 1:500, survivin (71G4B7) 1:1000 and p-S6K1 T389 (108D2) 1:1000, p-TSC2 S939 (rabbit polyclonal) 1:1000 obtained from Cell Signaling Technology.. Bcl-2 (N-19) 1:500, cyclin D1 (M-20) 1:500, p27Kip1 (C-19) 1:1000, cyclin A (C-19) 1:500, CDK4 (C-22) 1:1000, CDK6 (H-96) 1:500, and E2F1 (KH95) 1:500 were fom Santa Cruz. α-tubulin (CP06) 1:3000 was purchased from Calbiochem and GAPDH (rabbit polyclonal) 1:10000 from Trevigen.Bcl-2 (N-19) 1:500, cyclin D1 (M-20) 1:500, p27Kip1 (C-19) 1:1000, cyclin A (C-19) 1:500, CDK4 (C-22) 1:1000, CDK6 (H-96) 1:500, and E2F1 (KH95) 1:500 were fom Santa Cruz. α-tubulin (CP06) 1:3000 was purchased..

    Article Title: TSC1 promotes i NKT cell anergy and inhibits i NKT cell-mediated anti-tumor immunity
    Article Snippet: Fluorescence-conjugated anti-mouse TCRβ (H57-597), NK1.1 (PK136), CD44(IM7), B220(RA3-6B2), Gr-1(RB6-8C5), CD11c(N418), CD11b(M1/70), CD4 (GK1.5), CD8 (53-6.7), IFN-γ (XMG1.2), IL-4 (11B11), PD-1(RMP1-30), and ICOS(D10.G4.1) antibodies were purchased from BioLegend.Fluorescence-conjugated anti-mouse TCRβ (H57-597), NK1.1 (PK136), CD44(IM7), B220(RA3-6B2), Gr-1(RB6-8C5), CD11c(N418), CD11b(M1/70), CD4 (GK1.5), CD8 (53-6.7), IFN-γ (XMG1.2), IL-4 (11B11), PD-1(RMP1-30), and ICOS(D10.G4.1) antibodies were purchased from BioLegend.. Anti–phospho-p44/42 mitogen-activated protein kinase T202/Y204 (197G2), anti–phospho-S6 S240/244 (D68F8), anti–phospho-4E-BP1 T37/46 (236B4), anti–phospho-Akt S473 (193H12), TSC1 (D43E2), and TSC2 (D93F12) were purchased from Cell Signaling Technology, Inc. Anti–β-actin antibody was purchased from Sigma-Aldrich.. Cell surface staining was performed with 2% FBS-PBS.Cell surface staining was performed with 2% FBS-PBS.

    Article Title: TSC1 promotes i NKT cell anergy and inhibits i NKT cell-mediated anti-tumor immunity
    Article Snippet: CD11b(M1/70), CD4 (GK1.5), CD8 (53-6.7), IFN-γ (XMG1.2), IL-4 (11B11), PD-1(RMP1-30), and ICOS(D10.G4.1) antibodies were purchased from BioLegend. .. Anti–phospho-p44/42 mitogen-activated protein kinase T202/Y204 (197G2), anti–phospho-S6 S240/244 (D68F8), anti–phospho-4E-BP1 T37/46 (236B4), anti–phospho-Akt S473 (193H12), TSC1 (D43E2), and TSC2 (D93F12) were purchased from Cell Signaling Technology, Inc. Anti–β-actin antibody was purchased from Sigma-Aldrich. .. Cell surface staining was performed with 2% FBS-PBS.



    Similar Products

    tuberin tsc2 d93f12 xp rabbit mab  (Cell Signaling Technology Inc)
    86
    Cell Signaling Technology Inc tuberin tsc2 d93f12 xp rabbit mab
    Tuberin Tsc2 D93f12 Xp Rabbit Mab, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/tsc2+d93f12/pmc13068069-4-0-6
    Average 86 stars, based on 1 article reviews
    tuberin tsc2 d93f12 xp rabbit mab - by Bioz Stars, 2026-09
    86/100 stars
    		  Buy from Supplier
    tuberin tsc2 d93f12 xp  (Cell Signaling Technology Inc)
    96
    Cell Signaling Technology Inc tuberin tsc2 d93f12 xp
    Tuberin Tsc2 D93f12 Xp, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/tsc2+d93f12/Tuberin%2FTSC2+XP+Rabbit+mAb/bio_rxiv__2025__11__12__688066-187-49-52
    Average 96 stars, based on 1 article reviews
    tuberin tsc2 d93f12 xp - by Bioz Stars, 2026-09
    96/100 stars
    		  Buy from Supplier
    rabbit monoclonal ant tsc2 d93f12  (Cell Signaling Technology Inc)
    86
    Cell Signaling Technology Inc rabbit monoclonal ant tsc2 d93f12
    Rabbit Monoclonal Ant Tsc2 D93f12, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/tsc2+d93f12/pmc12588189-12-0-5
    Average 86 stars, based on 1 article reviews
    rabbit monoclonal ant tsc2 d93f12 - by Bioz Stars, 2026-09
    86/100 stars
    		  Buy from Supplier
    anti tsc2 rabbit monoclonal d93f12  (Cell Signaling Technology Inc)
    96
    Cell Signaling Technology Inc anti tsc2 rabbit monoclonal d93f12
    Anti Tsc2 Rabbit Monoclonal D93f12, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/tsc2+d93f12/Tuberin%2FTSC2+XP+Rabbit+mAb/pm40897470-247-30-35
    Average 96 stars, based on 1 article reviews
    anti tsc2 rabbit monoclonal d93f12 - by Bioz Stars, 2026-09
    96/100 stars
    		  Buy from Supplier
    anti-tuberin/tsc2 d93f12 xp #4308  (Cell Signaling Technology Inc)
    90
    Cell Signaling Technology Inc anti-tuberin/tsc2 d93f12 xp #4308
    Anti Tuberin/Tsc2 D93f12 Xp #4308, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/tsc2+d93f12/anti+tsc2/pm39892470-54-22-7
    Average 90 stars, based on 1 article reviews
    anti-tuberin/tsc2 d93f12 xp #4308 - by Bioz Stars, 2026-09
    90/100 stars
    		  Buy from Supplier
    tsc2 (d93f12, cell signaling #4308) antibody  (Cell Signaling Technology Inc)
    90
    Cell Signaling Technology Inc tsc2 (d93f12, cell signaling #4308) antibody
    Tsc2 (D93f12, Cell Signaling #4308) Antibody, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/tsc2+d93f12/anti+tsc2/pm39716490-179-1-4
    Average 90 stars, based on 1 article reviews
    tsc2 (d93f12, cell signaling #4308) antibody - by Bioz Stars, 2026-09
    90/100 stars
    		  Buy from Supplier

    Image Search Results