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anti p tfeb 87932s  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc anti p tfeb 87932s
    Anti P Tfeb 87932s, 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
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    Article Title: ESCRT-I fuels lysosomal degradation to restrict TFEB/TFE3 signaling via the Rag-mTORC1 pathway
    Article Snippet: The following antibodies were used: anti-Tsg101 (Cat. no. ab83) and anti-Vps28 (Cat. no. ab167172) from Abcam; anti-TFEB (Cat. no. 4240S), anti-P-TFEB (S122, Cat. no. 86843 and S211, Cat. no. 37681), anti-TFE3 (Cat. no. 14779S), anti-P-Ulk1 (S757, Cat. no. 6888), anti-P-4E-BP1 (Thr37/46, Cat. no. 2855), anti-P-S6K1 (Thr389, Cat. no. 9234), anti-mTOR (Cat. no. 2983), and anti-HA-Tag (Cat. no. 2367) from Cell Signaling Technologies; anti-LAMP1 (Cat. no. H4A3) from DSHB; anti-mono- and -polyubiquitinylated conjugates (Cat. no. BML-PW8810) from Enzo Life Sciences; anti-GFP (Cat. no. AF4240) from R&D Systems; anti-GAPDH (Cat. no. sc-25778) and anti-lamin A/C (Cat. no. sc-7292) from Santa Cruz; anti-histone H3 (Cat. no. H0164), anti-LAMP1 (Cat. no. L-1418), anti-vinculin (Cat. no. V9131-.2ML), and anti-tubulin (Cat. no. T5168) from Sigma-Aldrich; secondary HRP-conjugated goat anti-mouse, goat anti-rabbit, and bovine anti-goat antibodies from Jackson ImmunoResearch; and secondary Alexa Fluor 488–conjugated donkey anti-mouse and Alexa Fluor 647–conjugated donkey anti-rabbit antibodies from Thermo Fisher Scientific.

    Article Title: Toosendanin Induces Hepatocyte Damage by Inhibiting Autophagic Flux via TFEB-Mediated Lysosomal Dysfunction
    Article Snippet: Anti-LAMP1 (#15665), anti-CTSB (#31718), anti-TFEB (#37785), anti-p-TFEB(Ser211) (#37681), and anti-Histone H3(#4499) primary antibodies were purchased from Cell Signaling Technology (Billerica, CA, USA).

    Article Title: Identification of active natural products that induce lysosomal biogenesis by lysosome-based screening and biological evaluation
    Article Snippet: The following antibodies were used in this study: anti-actin (Proteintech, 66009-1-Ig), anti-LC3 (Abcam, ab48394), anti-s6k (Proteintech, 14485-1-AP), anti-p-s6k (Cell Signaling Technology, #9205), anti-TFEB (Proteintech, 13372-1-AP), and anti-p-TFEB (Cell Signaling Technology, #37681).

    Article Title: ESCRT III-mediated lysosomal repair improve renal tubular cell injury in cisplatin-induced AKI
    Article Snippet: The following antibodies were used in the experiments: anti-LC3B (Cell Signaling Technology, 2775, 12741), anti-GABARAPL1 (Cell Signaling Technology 13,733) anti-SQSTM1/p62 (Sigma-Aldrich, P0067), anti-LAMP1 (Santa Cruz Biotechnology, sc-18,821, sc-20,011), anti-CTSB/cathepsin B (Cell Signaling Technology 31,718), anti-NBR1 (Cell Signaling Technology 20,145), anti-BAX (proteintech 50,599–2-Ig), anti-BCL2 (Cell Signaling Technology, 3498), anti-cleaved CASP3/caspase 3 (Cell Signaling Technology, 9661), anti-EIF4EBP1 (Cell Signaling Technology, 9644), anti-p-EIF4EBP1 (Cell Signaling Technology, 2855), anti-RPS6KB1 (Cell Signaling Technology 34,475), anti-p-RPS6KB1(Cell Signaling Technology, 9209), anti-LGALS3/galectin 3 (Cell Signaling Technology 89,572), anti-TFEB (Cell Signaling Technology 83,010), anti-p-TFEB (Cell Signaling Technology 37,681), anti-GFP (Cell Signaling Technology, 2956), anti-Flag (Cell Signaling Technology, 14793S), anti-CHMP2A (proteintech 10,477–1-AP), anti-CHMP2B (proteintech 12,527–1-AP), anti-CHMP4B (proteintech 13,683–1-AP), anti-CHMP6 (proteintech 16,278–1-AP), anti-AQP1/aquaporin 1(abcam, ab168387).

    Article Title: ESCRT III-mediated lysosomal repair improve renal tubular cell injury in cisplatin-induced AKI.
    Article Snippet: The following antibodies were used in the experiments: anti-LC3B (Cell Signaling Technology, 2775, 12741), anti-GABARAPL1 (Cell Signaling Technology 13,733) anti-SQSTM1/p62 (Sigma-Aldrich, P0067), anti-LAMP1 (Santa Cruz Biotechnology, sc-18,821, sc-20,011), anti-CTSB/ cathepsin B (Cell Signaling Technology 31,718), anti-NBR1 (Cell Signaling Technology 20,145), anti-BAX (proteintech 50,599–2-Ig), anti-BCL2 (Cell Signaling Technology, 3498), anti-cleaved CASP3/caspase 3 (Cell Signaling Technology, 9661), anti-EIF4EBP1 (Cell Signaling Technology, 9644), anti-p-EIF4EBP1 (Cell Signaling Technology, 2855), antiRPS6KB1 (Cell Signaling Technology 34,475), anti-pRPS6KB1(Cell Signaling Technology, 9209), anti-LGALS3/ galectin 3 (Cell Signaling Technology 89,572), anti-TFEB (Cell Signaling Technology 83,010), anti-p-TFEB (Cell Signaling Technology 37,681), anti-GFP (Cell Signaling Technology, 2956), anti-Flag (Cell Signaling Technology, 14793S), anti-CHMP2A (proteintech 10,477–1-AP), antiCHMP2B (proteintech 12,527–1-AP), anti-CHMP4B (proteintech 13,683–1-AP), anti-CHMP6 (proteintech 16,278–1- AP), anti-AQP1/aquaporin 1(abcam, ab168387).

    Western Blot:

    Article Title: Sesquiterpenes from Carpesium faberi triggered ROS-induced apoptosis and protective autophagy in hepatocellular carcinoma cells.
    Article Snippet: Ten previously undescribed sesquiterpenes, carpespenes A–J (1–10), and eight known compounds (11–18), were isolated from the whole plants of Carpesium faberi.. Their structures were established by extensive analysis of HRESIMS, NMR, and ECD spectra.. Carpespene A (1) is eudesmanolide-type sesquiterpene lactone with an open five membered ring involving C-2 and C-3.

    Article Title: Quercetin ameliorates doxorubicin-induced atrial fibrillation via EGFR-mediated restoration of autophagic flux.
    Article Snippet: Background: Doxorubicin (DOX) is a widely used chemotherapeutic agent associated with significant cardiotoxicity, particularly manifesting as atrial fibrillation (AF), which adversely impacts patient quality of life.. Purpose: This study aimed to investigate the effects of quercetin and its underlying mechanisms in doxorubicin

    Article Title: Reducing FASN expression sensitizes acute myeloid leukemia cells to differentiation therapy
    Article Snippet: 293 T cells were maintained in DMEM (Sigma-Aldrich, St. Louis, MO, USA), supplemented with 5% FBS, 1% penicillin/streptomycin, and 1% Hepes (Sigma-Aldrich, Switzerland), and kept in a 7.5%CO 2 -95% air humidified atmosphere at 37 °C. .. Antibodies used were anti-FASN (3180; Cell Signaling, Switzerland), anti-LC3B (WB: NB600-1384, Novus biological, Switzerland; IF: 3868; Cell Signaling, Switzerland) anti-LAMP1 (14-1079-80; Thermofisher, Switzerland), anti-p62 (HPA003196; Sigma-Aldrich, Switzerland), anti-TFEB (4240; Cell Signaling, Switzerland), anti-p-TFEB (Ser 211) (37681S, Cell Signaling, Switzerland), anti-p-AKT (Ser473) (4060S, Cell Signaling, Switzerland) anti-PTEN (9552, Cell Signaling, Switzerland), anti-ULK1 (4776; Cell Signaling, Switzerland), anti-p-ULK1 (Ser757, equivalent to Ser758 of human ULK1) (6888; Cell Signaling, Switzerland), anti-ATG13 (6940; Cell Signaling, Switzerland), anti-pATG13 (Ser318) (600-401-C49; Rockland, Switzerland), anti p-mTOR (Ser2448) (5536; Cell Signaling, Switzerland), p4E-BP1 (Thr37/46) (2855; Cell Signaling, Switzerland), anti-α-tubulin (3873; Cell Signaling, Switzerland), anti-cleaved PARP (9541; Cell Signaling, Switzerland), anti yH2AX (2577; Cell Signaling, Switzerland) and anti-CD11b-PE (R0841; Dako, Switzerland). .. Whole-cell extracts were prepared using UREA lysis buffer and 30–60 μg of total protein was loaded on a 7.5% or 12% denaturing polyacrylamide self-cast gels (Bio-Rad, Switzerland).

    Cell-Signaling:

    Article Title: Sesquiterpenes from Carpesium faberi triggered ROS-induced apoptosis and protective autophagy in hepatocellular carcinoma cells.
    Article Snippet: Ten previously undescribed sesquiterpenes, carpespenes A–J (1–10), and eight known compounds (11–18), were isolated from the whole plants of Carpesium faberi.. Their structures were established by extensive analysis of HRESIMS, NMR, and ECD spectra.. Carpespene A (1) is eudesmanolide-type sesquiterpene lactone with an open five membered ring involving C-2 and C-3.



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    Image Search Results


    CuMn-ZIF nanozymes mitigate oxidative stress and protect chondrocytes in vitro. (A) Schematic diagram illustrating the proposed mechanism of CuMn-ZIF nanozymes in treating OA. CuMn-ZIF scavenges ROS, inhibits the phosphorylation of the PI3K–AKT–mTOR pathway, promotes TFEB nuclear translocation, enhances autophagic flux, and ultimately restores the balance between anabolism and catabolism in chondrocytes. (B) Intracellular ROS levels were measured using the DCFH-DA probe and (C) analyzed by flow cytometry following treatment with PBS, H 2 O 2 , H 2 O 2 + ZIF, or H 2 O 2 + CuMn-ZIF. (D) Representative fluorescence images and (E) quantitative analysis of mitochondrial membrane potential assessed by JC-1 staining (scale bar, 75 μm). Red fluorescence indicates JC-1 aggregates (healthy mitochondria), and green fluorescence indicates JC-1 monomers (depolarized mitochondria). (F) Representative WB images and (G) corresponding quantitative analysis of the protein expression levels of catabolic markers (ADAMTS-5, MMP-13) and anabolic markers (COL2A1, AGGRECAN). (H) Cell viability was determined by CCK-8 assay after treatment with PBS, H 2 O 2 , H 2 O 2 + ZIF, or H 2 O 2 + CuMn-ZIF. (I) Live/dead cell staining images (scale bar, 250 μm) using calcein-AM (green, live cells) and PI (red, dead cells), and the corresponding quantitative analysis of (J) calcein-AM and (K) PI fluorescence intensity. Statistical analysis was conducted using one-way ANOVA analysis. Data were represented as mean ± SD ( n = 3). **** P < 0.0001, *** P < 0.001, ** P < 0.01, * P < 0.05, ns: not significant.

    Journal: Biomaterials Research

    Article Title: Bimetallic Copper–Manganese Zeolitic Imidazolate Framework Nanozyme Scavenges Reactive Oxygen Species to Alleviate Osteoarthritis via Phosphoinositide 3-Kinase/Mammalian Target of Rapamycin Axis and Autophagic Flux Restoration

    doi: 10.34133/bmr.0306

    Figure Lengend Snippet: CuMn-ZIF nanozymes mitigate oxidative stress and protect chondrocytes in vitro. (A) Schematic diagram illustrating the proposed mechanism of CuMn-ZIF nanozymes in treating OA. CuMn-ZIF scavenges ROS, inhibits the phosphorylation of the PI3K–AKT–mTOR pathway, promotes TFEB nuclear translocation, enhances autophagic flux, and ultimately restores the balance between anabolism and catabolism in chondrocytes. (B) Intracellular ROS levels were measured using the DCFH-DA probe and (C) analyzed by flow cytometry following treatment with PBS, H 2 O 2 , H 2 O 2 + ZIF, or H 2 O 2 + CuMn-ZIF. (D) Representative fluorescence images and (E) quantitative analysis of mitochondrial membrane potential assessed by JC-1 staining (scale bar, 75 μm). Red fluorescence indicates JC-1 aggregates (healthy mitochondria), and green fluorescence indicates JC-1 monomers (depolarized mitochondria). (F) Representative WB images and (G) corresponding quantitative analysis of the protein expression levels of catabolic markers (ADAMTS-5, MMP-13) and anabolic markers (COL2A1, AGGRECAN). (H) Cell viability was determined by CCK-8 assay after treatment with PBS, H 2 O 2 , H 2 O 2 + ZIF, or H 2 O 2 + CuMn-ZIF. (I) Live/dead cell staining images (scale bar, 250 μm) using calcein-AM (green, live cells) and PI (red, dead cells), and the corresponding quantitative analysis of (J) calcein-AM and (K) PI fluorescence intensity. Statistical analysis was conducted using one-way ANOVA analysis. Data were represented as mean ± SD ( n = 3). **** P < 0.0001, *** P < 0.001, ** P < 0.01, * P < 0.05, ns: not significant.

    Article Snippet: For protein separation, electrophoresis was conducted at 90 V and the voltage was increased to 150 V. The proteins were transferred onto a nitrocellulose (NC) membrane and treated with blocking solution for 2 h, followed by overnight incubation with primary antibodies at 4 °C : AKT (CST, 9272), p-AKT (CST, 4060S), mTOR (CST, 2983), p-mTOR (CST, 5536), PI3K (Abclonal, A0982), p-PI3K (Abbkine, ABP50495 ), LC3B (Abcam, ab192890), P62 (Abcam, ab56416), TFEB (CST, 37681), and p-TFEB. glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (Absin, abs830030; Affinit, AF7021) was used as the housekeeping protein.

    Techniques: In Vitro, Phospho-proteomics, Translocation Assay, Flow Cytometry, Fluorescence, Membrane, Staining, Expressing, CCK-8 Assay

    CuMn-ZIF nanozymes enhance autophagic flux and promote TFEB nuclear translocation in chondrocytes in vitro. (A) Representative WB images and (B) quantitative analysis of autophagy-related proteins (LC3-II/I and P62) in chondrocytes treated with PBS, H 2 O 2 , H 2 O 2 + ZIF, or H 2 O 2 + CuMn-ZIF. (C) Representative confocal images and (D) quantitative analysis of autophagic flux using the mCherry-GFP-LC3 fluorescent probe. Yellow puncta (mCherry + GFP + ) represent autophagosomes, and red puncta (mCherry + GFP − ) represent autolysosomes (scale bar, 250 μm). (E) Immunofluorescence images showing the subcellular localization of TFEB (FITC, green) with nuclei counterstained by DAPI (blue), and (F) the corresponding statistical analysis of nuclear-to-cytoplasmic TFEB fluorescence intensity ratio (scale bar, 75 μm). (G) Representative WB images of p-TFEB and total TFEB and (H) quantitative analysis of p-TFEB/TFEB protein levels. (I) Flow cytometry analysis and (J) quantification of lysosomal pH using LysoTracker Red staining. (K) Representative confocal images and (L) quantitative analysis of autophagic flux using the mCherry-GFP-LC3 fluorescent probe according to siNC and siTFEB treatment (scale bar, 100 μm). Statistical analysis was conducted using one-way ANOVA analysis. Data were represented as mean ± SD ( n = 3). **** P < 0.0001, *** P < 0.001, ** P < 0.01, * P < 0.05, ns: not significant.

    Journal: Biomaterials Research

    Article Title: Bimetallic Copper–Manganese Zeolitic Imidazolate Framework Nanozyme Scavenges Reactive Oxygen Species to Alleviate Osteoarthritis via Phosphoinositide 3-Kinase/Mammalian Target of Rapamycin Axis and Autophagic Flux Restoration

    doi: 10.34133/bmr.0306

    Figure Lengend Snippet: CuMn-ZIF nanozymes enhance autophagic flux and promote TFEB nuclear translocation in chondrocytes in vitro. (A) Representative WB images and (B) quantitative analysis of autophagy-related proteins (LC3-II/I and P62) in chondrocytes treated with PBS, H 2 O 2 , H 2 O 2 + ZIF, or H 2 O 2 + CuMn-ZIF. (C) Representative confocal images and (D) quantitative analysis of autophagic flux using the mCherry-GFP-LC3 fluorescent probe. Yellow puncta (mCherry + GFP + ) represent autophagosomes, and red puncta (mCherry + GFP − ) represent autolysosomes (scale bar, 250 μm). (E) Immunofluorescence images showing the subcellular localization of TFEB (FITC, green) with nuclei counterstained by DAPI (blue), and (F) the corresponding statistical analysis of nuclear-to-cytoplasmic TFEB fluorescence intensity ratio (scale bar, 75 μm). (G) Representative WB images of p-TFEB and total TFEB and (H) quantitative analysis of p-TFEB/TFEB protein levels. (I) Flow cytometry analysis and (J) quantification of lysosomal pH using LysoTracker Red staining. (K) Representative confocal images and (L) quantitative analysis of autophagic flux using the mCherry-GFP-LC3 fluorescent probe according to siNC and siTFEB treatment (scale bar, 100 μm). Statistical analysis was conducted using one-way ANOVA analysis. Data were represented as mean ± SD ( n = 3). **** P < 0.0001, *** P < 0.001, ** P < 0.01, * P < 0.05, ns: not significant.

    Article Snippet: For protein separation, electrophoresis was conducted at 90 V and the voltage was increased to 150 V. The proteins were transferred onto a nitrocellulose (NC) membrane and treated with blocking solution for 2 h, followed by overnight incubation with primary antibodies at 4 °C : AKT (CST, 9272), p-AKT (CST, 4060S), mTOR (CST, 2983), p-mTOR (CST, 5536), PI3K (Abclonal, A0982), p-PI3K (Abbkine, ABP50495 ), LC3B (Abcam, ab192890), P62 (Abcam, ab56416), TFEB (CST, 37681), and p-TFEB. glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (Absin, abs830030; Affinit, AF7021) was used as the housekeeping protein.

    Techniques: Translocation Assay, In Vitro, Immunofluorescence, Fluorescence, Flow Cytometry, Staining