Virus:Article Title: LKB1 controls inflammatory potential through CRTC2-dependent histone acetylation.
Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Rabbit monoclonal anti-human/mouse beta-actin Cell Signaling Cat4970; RRID: AB_2223172 Rabbit monoclonal anti-Lamin B1 Cell Signaling Cat#12586; RRID: AB_2650517 Rabbit polyclonal anti-alpha-tubulin Cell Signaling Cat#2144; RRID: AB_2210548 Rabbit monoclonal anti-LKB1 Cell Signaling Cat#3047; RRID: AB_2198327 Rabbit polyclonal anti-CRTC2 Proteintech Cat#12497-1-AP; RRID: AB_2260910 Rabbit polyclonal anti-CREB1 Proteintech Cat#12208-1-AP; RRID: AB_2245417 Rabbit monoclonal anti-phospho-CREB (Ser133) Cell Signaling Cat#9198; RRID: AB_2561044 Rabbit monoclonal anti-phospho-p70 S6 kinase (Thr389) Cell Signaling Cat#97596; RRID: AB_2800283 Rabbit monoclonal anti-p70 S6 kinase Cell Signaling Cat#2708; RRID: AB_390722 Rabbit polyclonal anti-phospho-S6 ribosomal protein (Ser235/236) Cell Signaling Cat#2211; RRID: AB_331679 Rabbit monoclonal anti-S6 ribosomal protein Cell Signaling Cat#2217; RRID: AB_331355 Rabbit monoclonal anti-phospho-4EBP1 (Thr37/46) Cell Signaling Cat#2855; RRID: AB_560835 Rabbit monoclonal anti-phospho-HDAC4 (Ser246)/HDAC5 (Ser259)/HDAC7 (Ser155) Cell Signaling Cat#3443; RRID: AB_2118723 Rabbit polyclonal anti-phospho-HDAC4 (Ser246)/HDAC5 (Ser259)/HDAC9 (Ser220) Invitrogen Cat#PA5-37835; RRID: AB_2554443 Rabbit monoclonal anti-HDAC4 Cell Signaling Cat#15164; RRID: AB_2798733 Rabbit monoclonal anti-HDAC5 Cell Signaling Cat#20458; RRID: AB_2713973 Rabbit monoclonal anti-HDAC7 Cell Signaling Cat#33418; RRID: AB_2716756 Rabbit polyclonal anti-HDAC9 Invitrogen Cat#PA5-90264; RRID: AB_2805993 Goat anti-rabbit IgG, HRP-linked Cell Signaling Cat#7074; RRID: AB_2099233 Histone H3K27ac Active Motif Cat#39133; RRID: AB_2561016 Normal Rabbit IgG Cell Signaling Cat#2729; RRID: AB_1031062 Bacterial and Virus Strains One Shot Stbl3 Chemically Competent E. coli Invitrogen Cat#C737303 Chemicals, Peptides, and Recombinant Proteins DMEM Wisent Bioproducts Cat#319-015-CL RPMI Medium 1640 Gibco Cat#11875-093 Opti-MEM I Gibco Cat#31985070 HBSS Gibco Cat#14185-052 Nu-Serum IV Culture Supplement Corning Cat#355504 Penicillin-streptomycin Gibco Cat#15070-063 L-Glutamine Gibco Cat#25030-081 LPS from E. coli 0111:B4 Sigma-Aldrich Cat#L2630 Standard LPS, E. coli K12 Invivogen tlrl-eklps Recombinant murine IL-1b Peprotech Cat#211-11B Recombinant mouse IL-1a R&D Systems Cat#400-ML Recombinant mouse TNF-a R&D Systems Cat#410-MT (Continued on next page) Molecular Cell 83, 1872–1886.e1–e5, June 1, 2023 e1 .. REAGENT or RESOURCE SOURCE IDENTIFIER Recombinant mouse IFN-a BioLegend Cat#752802 Recombinant mouse IFN-b R&D Systems Cat#12410-1 Recombinant murine IFN-g Peprotech Cat#315-05 Rapamycin Millipore Sigma Cat#37094 A 485 Tocris Bioscience Cat#6387 Vorinostat Selleck Chemicals Cat#S1047 HG-9-91-01 MedChemExpress Cat#HY-15776 BbsI NEB Cat#R0539 Dynabeads Protein A Invitrogen Cat#10001D Proteinase K Invitrogen Cat#25530-015 Pierce 16% Formaldehyde (w/v), methanol-free Thermo Fisher Cat#28908 Critical Commercial Assays Qiagen RNAeasy Mini Kit Qiagen Cat#74104 QIAquick PCR Purification Kit Qiagen #28104 HiSpeed Plasmid Maxi Kit Qiagen #12662 Histone acetyltransferase activity assay kit Abcam #ab65352 Mouse IL-6 ELISA kit Thermo Fisher #88-7064-88 Mouse LIF Quantikine ELISA kit R&D Systems MLF00 Mouse Cytokine/Chemokine 44-Plex Discovery Assay Array Eve Technologies MD44 cOmplete, Mini, EDTA-free Protease Inhibitor Cocktail Roche 11836170001 PhosSTOP Roche 4906845001 Nuclear extraction kit Abcam ab113474 ON-TARGETplus Mouse Stk11 siRNA Horizon Discovery L-044342-00-0005 ON-TARGETplus Mouse Socs3 siRNA Horizon Discovery L-040626-01-0005 ON-TARGETplus Mouse Myd88 siRNA Horizon Discovery L-063057-00-0005 ON-TARGETplus Mouse Prkaa1 siRNA Horizon Discovery L-041035-00-0005 ON-TARGETplus Mouse Prkaa2 siRNA Horizon Discovery L-040809-00-0005 ON-TARGETplus Mouse Sik1 siRNA Horizon Discovery L-044399-00-0005 ON-TARGETplus Mouse Sik2 siRNA Horizon Discovery L-041008-00-0005 ON-TARGETplus Mouse Sik3 siRNA Horizon Discovery L-061908-00-0005 ON-TARGETplus Mouse Crtc1 siRNA Horizon Discovery L-066810-01-0005 ON-TARGETplus Mouse Crtc2 siRNA Horizon Discovery L-049303-00-0005 ON-TARGETplus Mouse Hdac4 siRNA Horizon Discovery L-043626-00-0005 ON-TARGETplus Mouse Hdac9 siRNA Horizon Discovery L-066143-00-0005 ON-TARGETplus Non-targeting Control Pool Horizon Discovery D-001810-10-05 DharmaFECT 1 Transfection Reagent Horizon Discovery Cat#T-2001 Deposited data RNA-sequencing and ChIPsequencing data This manuscript GEO: GSE214824 Experimental Models: Cell Lines Wild type mouse embryonic fibroblasts (MEFs) Faubert et al.64 N/A Lkb1-/- MEFs Faubert et al.64 N/A Ampk-/- MEFs Rabinovitch et al.81 N/A Lkb1/Crtc2-/- MEFs This manuscript N/A (Continued on next page) e2 Molecular Cell 83, 1872–1886.e1–e5, June 1, 2023
Article Title: Metabolic Profiling Using Stable Isotope Tracing Reveals Distinct Patterns of Glucose Utilization by Physiologically Activated CD8 + T Cells.
Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Monoclonal anti-mouse CD3, eBioscience Functional Grade, Purified Invitrogen Clone # 145-2C11; RRID: AB_468849 Monoclonal anti-mouse CD28, eBioscience Functional Grade, Purified Invitrogen Clone # 37.51; RRID: AB_468923 Rat monoclonal anti-mouse CD25 eBioscience Clone # PC61.5 Rat monoclonal anti-mouse CD4 eBioscience Clone # GK1.5 Rat monoclonal anti-mouse CD62L eBioscience Clone # MEL-14 Rat monoclonal anti-mouse IFN-gamma eBioscience Clone # XMG1.2 Rat monoclonal anti-mouse CD8 eBioscience Clone # 53-6.7 Mouse monoclonal anti-mouse CD45.2 eBioscience Clone # 104 Rat monoclonal anti-mouse CD127 BioLegend Clone # A7R34; RRID AB_11218800 Hamster monoclonal anti-mouse KLRG1 eBioscience Clone # 2F1; RRID AB_1311265 Rat monoclonal anti-mouse CD44 eBioscience Clone # IM7 Rabbit polyclonal anti-human/mouse beta-actin Cell Signaling Cat # 4967; RRID AB_330288 Mouse monoclonal anti-mouse CD90.1 (Thy-1.1) eBioscience Clone # HIS51 Mouse monoclonal anti-mouse CD90.2 (Thy1-1.2) eBioscience Clone # 30-H12 Rabbit polyclonal anti mouse PHGDH Antibody ThermoFisher Scientific Cat # PA5-27578; RRID: AB_2545054 H-2Kb/OVA257 Tetramer Baylor College of Medicine N/A Bacterial and Virus Strains Listeria monocytogenes rLM-OVA van der Windt et al., 2012 N/A Luciferase shRNA (LMPd-Amt backbone): AGCTCCCGTGAATT GGAATCCTAGTGAAGCCACAGATGTAGGATTCCAATTCAGCG GGAGCC Chen et al., 2014 for vector backbone N/A Phgdh shRNA (LMPd-Amt backbone): tgctgttgacagtgagcgcgcact ggatgtgtttacagaatagtgaagccacagatgtattctgtaaacacatccagtgcatg cctactgcctcgga This paper N/A Chemicals, Peptides, and Recombinant Proteins [U-13C]-glucose Cambridge Isotopes Cat # CLM-1396 IMDM GlutaMAX Thermo Fisher Cat # 3198-022 Heat-inactivated FCS Biochrom Cat # 0115 – Lot# 1289W Penicillin-streptomycin Millipore Cat # A2213 2-mercaptoethanol GIBCO Cat # 21985-023 recombinant murine IL-2 Peprotech Cat # 212-12 recombinant murine IL-7 Peprotech Cat # 217-17 Phorbol 12-myristate 13-acetate (PMA) Sigma-Aldrich Cat # P-8139 Ionomycin Sigma-Aldrich Cat # 10634 FCCP Cayman Cat # 15218 Oligomycin Sigma-Aldrich Cat # 1404-19-9 Rotenone Cayman Cat # 13955 Antimycin A Sigma Aldrich Cat # A8674 XF Base Medium Minimal DMEM without Phenol Red Agilent Cat # 103335-100 Glucose R 99,5% D(+) Roth Cat # HN06.1 L-Glutamine Biochrom AG Cat # M11-004 Sodium Pyruvate GIBCO Cat # 11360-070 Poly-D-lysine hydrobromide Sigma Aldrich Cat # P6407 (Continued on next page) e1 Immunity 51, 1–15.e1–e5, November 19, 2019 ..
Recombinant:Article Title: LKB1 controls inflammatory potential through CRTC2-dependent histone acetylation.
Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Rabbit monoclonal anti-human/mouse beta-actin Cell Signaling Cat4970; RRID: AB_2223172 Rabbit monoclonal anti-Lamin B1 Cell Signaling Cat#12586; RRID: AB_2650517 Rabbit polyclonal anti-alpha-tubulin Cell Signaling Cat#2144; RRID: AB_2210548 Rabbit monoclonal anti-LKB1 Cell Signaling Cat#3047; RRID: AB_2198327 Rabbit polyclonal anti-CRTC2 Proteintech Cat#12497-1-AP; RRID: AB_2260910 Rabbit polyclonal anti-CREB1 Proteintech Cat#12208-1-AP; RRID: AB_2245417 Rabbit monoclonal anti-phospho-CREB (Ser133) Cell Signaling Cat#9198; RRID: AB_2561044 Rabbit monoclonal anti-phospho-p70 S6 kinase (Thr389) Cell Signaling Cat#97596; RRID: AB_2800283 Rabbit monoclonal anti-p70 S6 kinase Cell Signaling Cat#2708; RRID: AB_390722 Rabbit polyclonal anti-phospho-S6 ribosomal protein (Ser235/236) Cell Signaling Cat#2211; RRID: AB_331679 Rabbit monoclonal anti-S6 ribosomal protein Cell Signaling Cat#2217; RRID: AB_331355 Rabbit monoclonal anti-phospho-4EBP1 (Thr37/46) Cell Signaling Cat#2855; RRID: AB_560835 Rabbit monoclonal anti-phospho-HDAC4 (Ser246)/HDAC5 (Ser259)/HDAC7 (Ser155) Cell Signaling Cat#3443; RRID: AB_2118723 Rabbit polyclonal anti-phospho-HDAC4 (Ser246)/HDAC5 (Ser259)/HDAC9 (Ser220) Invitrogen Cat#PA5-37835; RRID: AB_2554443 Rabbit monoclonal anti-HDAC4 Cell Signaling Cat#15164; RRID: AB_2798733 Rabbit monoclonal anti-HDAC5 Cell Signaling Cat#20458; RRID: AB_2713973 Rabbit monoclonal anti-HDAC7 Cell Signaling Cat#33418; RRID: AB_2716756 Rabbit polyclonal anti-HDAC9 Invitrogen Cat#PA5-90264; RRID: AB_2805993 Goat anti-rabbit IgG, HRP-linked Cell Signaling Cat#7074; RRID: AB_2099233 Histone H3K27ac Active Motif Cat#39133; RRID: AB_2561016 Normal Rabbit IgG Cell Signaling Cat#2729; RRID: AB_1031062 Bacterial and Virus Strains One Shot Stbl3 Chemically Competent E. coli Invitrogen Cat#C737303 Chemicals, Peptides, and Recombinant Proteins DMEM Wisent Bioproducts Cat#319-015-CL RPMI Medium 1640 Gibco Cat#11875-093 Opti-MEM I Gibco Cat#31985070 HBSS Gibco Cat#14185-052 Nu-Serum IV Culture Supplement Corning Cat#355504 Penicillin-streptomycin Gibco Cat#15070-063 L-Glutamine Gibco Cat#25030-081 LPS from E. coli 0111:B4 Sigma-Aldrich Cat#L2630 Standard LPS, E. coli K12 Invivogen tlrl-eklps Recombinant murine IL-1b Peprotech Cat#211-11B Recombinant mouse IL-1a R&D Systems Cat#400-ML Recombinant mouse TNF-a R&D Systems Cat#410-MT (Continued on next page) Molecular Cell 83, 1872–1886.e1–e5, June 1, 2023 e1 .. REAGENT or RESOURCE SOURCE IDENTIFIER Recombinant mouse IFN-a BioLegend Cat#752802 Recombinant mouse IFN-b R&D Systems Cat#12410-1 Recombinant murine IFN-g Peprotech Cat#315-05 Rapamycin Millipore Sigma Cat#37094 A 485 Tocris Bioscience Cat#6387 Vorinostat Selleck Chemicals Cat#S1047 HG-9-91-01 MedChemExpress Cat#HY-15776 BbsI NEB Cat#R0539 Dynabeads Protein A Invitrogen Cat#10001D Proteinase K Invitrogen Cat#25530-015 Pierce 16% Formaldehyde (w/v), methanol-free Thermo Fisher Cat#28908 Critical Commercial Assays Qiagen RNAeasy Mini Kit Qiagen Cat#74104 QIAquick PCR Purification Kit Qiagen #28104 HiSpeed Plasmid Maxi Kit Qiagen #12662 Histone acetyltransferase activity assay kit Abcam #ab65352 Mouse IL-6 ELISA kit Thermo Fisher #88-7064-88 Mouse LIF Quantikine ELISA kit R&D Systems MLF00 Mouse Cytokine/Chemokine 44-Plex Discovery Assay Array Eve Technologies MD44 cOmplete, Mini, EDTA-free Protease Inhibitor Cocktail Roche 11836170001 PhosSTOP Roche 4906845001 Nuclear extraction kit Abcam ab113474 ON-TARGETplus Mouse Stk11 siRNA Horizon Discovery L-044342-00-0005 ON-TARGETplus Mouse Socs3 siRNA Horizon Discovery L-040626-01-0005 ON-TARGETplus Mouse Myd88 siRNA Horizon Discovery L-063057-00-0005 ON-TARGETplus Mouse Prkaa1 siRNA Horizon Discovery L-041035-00-0005 ON-TARGETplus Mouse Prkaa2 siRNA Horizon Discovery L-040809-00-0005 ON-TARGETplus Mouse Sik1 siRNA Horizon Discovery L-044399-00-0005 ON-TARGETplus Mouse Sik2 siRNA Horizon Discovery L-041008-00-0005 ON-TARGETplus Mouse Sik3 siRNA Horizon Discovery L-061908-00-0005 ON-TARGETplus Mouse Crtc1 siRNA Horizon Discovery L-066810-01-0005 ON-TARGETplus Mouse Crtc2 siRNA Horizon Discovery L-049303-00-0005 ON-TARGETplus Mouse Hdac4 siRNA Horizon Discovery L-043626-00-0005 ON-TARGETplus Mouse Hdac9 siRNA Horizon Discovery L-066143-00-0005 ON-TARGETplus Non-targeting Control Pool Horizon Discovery D-001810-10-05 DharmaFECT 1 Transfection Reagent Horizon Discovery Cat#T-2001 Deposited data RNA-sequencing and ChIPsequencing data This manuscript GEO: GSE214824 Experimental Models: Cell Lines Wild type mouse embryonic fibroblasts (MEFs) Faubert et al.64 N/A Lkb1-/- MEFs Faubert et al.64 N/A Ampk-/- MEFs Rabinovitch et al.81 N/A Lkb1/Crtc2-/- MEFs This manuscript N/A (Continued on next page) e2 Molecular Cell 83, 1872–1886.e1–e5, June 1, 2023
Article Title: Metabolic Profiling Using Stable Isotope Tracing Reveals Distinct Patterns of Glucose Utilization by Physiologically Activated CD8 + T Cells.
Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Monoclonal anti-mouse CD3, eBioscience Functional Grade, Purified Invitrogen Clone # 145-2C11; RRID: AB_468849 Monoclonal anti-mouse CD28, eBioscience Functional Grade, Purified Invitrogen Clone # 37.51; RRID: AB_468923 Rat monoclonal anti-mouse CD25 eBioscience Clone # PC61.5 Rat monoclonal anti-mouse CD4 eBioscience Clone # GK1.5 Rat monoclonal anti-mouse CD62L eBioscience Clone # MEL-14 Rat monoclonal anti-mouse IFN-gamma eBioscience Clone # XMG1.2 Rat monoclonal anti-mouse CD8 eBioscience Clone # 53-6.7 Mouse monoclonal anti-mouse CD45.2 eBioscience Clone # 104 Rat monoclonal anti-mouse CD127 BioLegend Clone # A7R34; RRID AB_11218800 Hamster monoclonal anti-mouse KLRG1 eBioscience Clone # 2F1; RRID AB_1311265 Rat monoclonal anti-mouse CD44 eBioscience Clone # IM7 Rabbit polyclonal anti-human/mouse beta-actin Cell Signaling Cat # 4967; RRID AB_330288 Mouse monoclonal anti-mouse CD90.1 (Thy-1.1) eBioscience Clone # HIS51 Mouse monoclonal anti-mouse CD90.2 (Thy1-1.2) eBioscience Clone # 30-H12 Rabbit polyclonal anti mouse PHGDH Antibody ThermoFisher Scientific Cat # PA5-27578; RRID: AB_2545054 H-2Kb/OVA257 Tetramer Baylor College of Medicine N/A Bacterial and Virus Strains Listeria monocytogenes rLM-OVA van der Windt et al., 2012 N/A Luciferase shRNA (LMPd-Amt backbone): AGCTCCCGTGAATT GGAATCCTAGTGAAGCCACAGATGTAGGATTCCAATTCAGCG GGAGCC Chen et al., 2014 for vector backbone N/A Phgdh shRNA (LMPd-Amt backbone): tgctgttgacagtgagcgcgcact ggatgtgtttacagaatagtgaagccacagatgtattctgtaaacacatccagtgcatg cctactgcctcgga This paper N/A Chemicals, Peptides, and Recombinant Proteins [U-13C]-glucose Cambridge Isotopes Cat # CLM-1396 IMDM GlutaMAX Thermo Fisher Cat # 3198-022 Heat-inactivated FCS Biochrom Cat # 0115 – Lot# 1289W Penicillin-streptomycin Millipore Cat # A2213 2-mercaptoethanol GIBCO Cat # 21985-023 recombinant murine IL-2 Peprotech Cat # 212-12 recombinant murine IL-7 Peprotech Cat # 217-17 Phorbol 12-myristate 13-acetate (PMA) Sigma-Aldrich Cat # P-8139 Ionomycin Sigma-Aldrich Cat # 10634 FCCP Cayman Cat # 15218 Oligomycin Sigma-Aldrich Cat # 1404-19-9 Rotenone Cayman Cat # 13955 Antimycin A Sigma Aldrich Cat # A8674 XF Base Medium Minimal DMEM without Phenol Red Agilent Cat # 103335-100 Glucose R 99,5% D(+) Roth Cat # HN06.1 L-Glutamine Biochrom AG Cat # M11-004 Sodium Pyruvate GIBCO Cat # 11360-070 Poly-D-lysine hydrobromide Sigma Aldrich Cat # P6407 (Continued on next page) e1 Immunity 51, 1–15.e1–e5, November 19, 2019 ..
Article Title: Ribosome-Targeting Antibiotics Impair T Cell Effector Function and Ameliorate Autoimmunity by Blocking Mitochondrial Protein Synthesis
Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Hamster monoclonal anti-mouse CD3 Bio X Cell Cat#: BE0001-1; RRID: AB_1107634 Hamster monoclonal anti-mouse CD28 Bio X Cell Cat#: BE00015-1; RRID: AB_1107624 Hamster monoclonal anti-mouse IFN-gamma Bio X Cell Cat#: BE0055; RRID: AB_1107694 Rat recombinant anti-mouse IL-12 p40 Bio X Cell Clone 17.8; RRID: AB_1107698 Rat monoclonal anti-mouse IL-4 Bio X Cell Cat#: BE0045; RRID: AB_1107707 Rat monoclonal anti-mouse CD4 eBioscience Clone 48-0041; RRID: AB_1107642 Rat monoclonal anti-mouse/rat IL-17A eBioscience Clone eBio17B7; RRID: AB_763580 Rat monoclonal anti-mouse IL-13 eBioscience Clone eBio13A; RRID: AB_763559 Rat monoclonal anti-mouse IFN-gamma eBioscience Clone XMG1.2; RRID: AB_315403 Mouse monoclonal anti-human/mouse T-bet eBioscience Clone eBio4B10; RRID: AB_763634 Rat monoclonal anti-mouse RORgt eBioscience Clone B2D; RRID: AB_315403 Rat monoclonal anti-mouse/human GATA-3 eBioscience Clone TWAJ; RRID: AB_1963600 Rat monoclonal anti-mouse/rat FoxP3 eBioscience Clone FJK-16s; RRID: AB_1518812 Mouse monoclonal anti-mouse CD154 (CD40 Ligand) eBioscience Clone MR1; RRID: AB_2561561 Mouse monoclonal anti-mouse/rat COX1 Abcam Cat#: ab154477 Mouse monoclonal anti-human CD4 Invitrogen Clone SK3; RRID: AB_1937227 Mouse monoclonal anti-human IL-17A eBIoscience Clone eBio64DEC17; RRID: AB_11063994 Mouse monoclonal anti-mouse antihuman CD3 BioLegend Ultra-LEAFTM Clone OKT3; RRID: AB_11150592 Mouse monoclonal anti-mouse antihuman CD28 BioLegend Ultra-LEAFTM Clone CD28.2; RRID: AB_314304 Recombinant anti-SDHA mAb Abcam aCat#: ab139181 Recombinant anti-ND1 mAb Abcam Cat#: ab181848 Recombinant anti-COX1 mAb Abcam Cat#: ab139181 HIF-1 alpha Rabbit mAb Novus biological Cat#: NB100-479: RRID: AB_10000633 AMPKa (23A3) Rabbit mAb Cell Signaling Cat#: 2603; RRID:AB_490795 Phospho-AMPKa (Thr172) Rabbit mAb Cell Signaling Cat#: 2535 Anti-rabbit IgG, HRP-linked Antibody Cell Signaling Cat#: 7074; RRID:AB_2099233 Anti-mouse IgG, HRP-linked Antibody Cell Signaling Cat#: 7076; RRID:AB_330924 Rabbit polyclonal anti-human/mouse beta-actin Cell Signaling Cat#: 4967; RRID:AB_330288 Chemicals, Peptides, and Recombinant Proteins IMDM GlutaMAX Thermo Fisher 3198-022 RPMI 1640 GlutaMAX Thermo Fisher 61870-010 X-Vivo 15 medium Lonza BE02-060F Seahorse XF DMEM medium, pH 7.4 Agilent 103575-100 Head-inactivated FCS Biochrom S 0115 – Lot# 1289W Penicillin-streptomycin Millipore P0781 2-mercaptoethanol Gibco #21985-023 Recombinant human TGF-beta Peprotech 100-21 (Continued on next page) e1 Immunity 54, 68–83.e1–e6, January 12, 2021 .. REAGENT or RESOURCE SOURCE IDENTIFIER Recombinant murine IL-6 Peprotech 211-11B Recombinant murine IL-1beta Peprotech 211-11B Recombinant murine IL-12 Peprotech 210-12 Recombinant murine IL-7 Peprotech 217-17 Recombinant murine IL-4 Preprotech 214-14 Recombinant human IL-12 p70 (HEK293 derived) Peprotech 200-12H Recombinant human IL-1beta R&D Systems 201-LB-005 Recombinant human IL-23 R&D Systems 1290-IL-010/CF Recombinant human IL-6 Peprotech 200-06 Recombinant human IL-21 Peprotech 200-21 Phorbol 12-myristate 13-acetate (PMA) Sigma-Aldrich P-8139 Ionomycin Sigma-Aldrich 10634 Tetramethzlyrhodamine, Ethyl Ester, Percholorate (TMRE) Invitrogen T669 Pierce RIPA Buffer Thermo Scientific 89900 Oligomycin Sigma Aldrich 75351 FCCP Cayman 15218 Rotenone Cayman 13955 Antimycin A Sigma Aldrich A8674 XF Base Medium Minimal DMEM without Phenol Red Agilent 103 Glucose R 99.5% D(+) Roth HN06.1 L-Glutamine Biochrom AG M11-004 Poly-D-lysine Hydrobromide Sigma Aldrich P6407 RNAse OUT Ribonuclease inhibitor (recombinant) Invitrogen #10777-019 CellTrace Violet Cell Proliferation Kit Life Technologies #65-0866-14 Fixable Viability Dye eFluor 780 eBioscience #65-0865-14 BD GolgiStop BD Biosciences #51-2092KZ FoxP3/transcription factor staining buffer set eBiosciences #00-5523-00 DMOG Cayman chemicals 71210 Lactate oxidase from Aerococcus viridans Sigma L9795-50UN Catalase, aspergillus niger Merck Millipore 219261-100KU Linezolid Chem-Impex 29723 Tigecycline hydrate Sigma PZ0021 Thiamphenicol Chem-Impex 29857 Vancomycin Cayman-Chemical #15327 FITC BrdU Flow Kit (RUO) BD 557891 DS18561882 inhibitor Medchemexpress HY-130251 Chaetocin from chaetomium minutum Merck C9492 DIF-1 Sigma SML0599 Mitomycin Cayman 11435 Paclitaxel Cayman 10461 SP600125 Inhibitor Cayman 10010466 ATP-polyamine-biotin Medchemexpress HY-D0183 Adenosine 5’triphosphate disodium salt hydrate Merck A26209 MCC905 InvivoGen inh-mcc (Continued on next page) ll OPEN ACCESSArticle Immunity 54, 68–83.e1–e6, January 12, 2021 e2
Article Title: Ribosome-Targeting Antibiotics Impair T Cell Effector Function and Ameliorate Autoimmunity by Blocking Mitochondrial Protein Synthesis.
Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Hamster monoclonal anti-mouse CD3 Bio X Cell Cat#: BE0001-1; RRID: AB_1107634 Hamster monoclonal anti-mouse CD28 Bio X Cell Cat#: BE00015-1; RRID: AB_1107624 Hamster monoclonal anti-mouse IFN-gamma Bio X Cell Cat#: BE0055; RRID: AB_1107694 Rat recombinant anti-mouse IL-12 p40 Bio X Cell Clone 17.8; RRID: AB_1107698 Rat monoclonal anti-mouse IL-4 Bio X Cell Cat#: BE0045; RRID: AB_1107707 Rat monoclonal anti-mouse CD4 eBioscience Clone 48-0041; RRID: AB_1107642 Rat monoclonal anti-mouse/rat IL-17A eBioscience Clone eBio17B7; RRID: AB_763580 Rat monoclonal anti-mouse IL-13 eBioscience Clone eBio13A; RRID: AB_763559 Rat monoclonal anti-mouse IFN-gamma eBioscience Clone XMG1.2; RRID: AB_315403 Mouse monoclonal anti-human/mouse T-bet eBioscience Clone eBio4B10; RRID: AB_763634 Rat monoclonal anti-mouse RORgt eBioscience Clone B2D; RRID: AB_315403 Rat monoclonal anti-mouse/human GATA-3 eBioscience Clone TWAJ; RRID: AB_1963600 Rat monoclonal anti-mouse/rat FoxP3 eBioscience Clone FJK-16s; RRID: AB_1518812 Mouse monoclonal anti-mouse CD154 (CD40 Ligand) eBioscience Clone MR1; RRID: AB_2561561 Mouse monoclonal anti-mouse/rat COX1 Abcam Cat#: ab154477 Mouse monoclonal anti-human CD4 Invitrogen Clone SK3; RRID: AB_1937227 Mouse monoclonal anti-human IL-17A eBIoscience Clone eBio64DEC17; RRID: AB_11063994 Mouse monoclonal anti-mouse antihuman CD3 BioLegend Ultra-LEAFTM Clone OKT3; RRID: AB_11150592 Mouse monoclonal anti-mouse antihuman CD28 BioLegend Ultra-LEAFTM Clone CD28.2; RRID: AB_314304 Recombinant anti-SDHA mAb Abcam aCat#: ab139181 Recombinant anti-ND1 mAb Abcam Cat#: ab181848 Recombinant anti-COX1 mAb Abcam Cat#: ab139181 HIF-1 alpha Rabbit mAb Novus biological Cat#: NB100-479: RRID: AB_10000633 AMPKa (23A3) Rabbit mAb Cell Signaling Cat#: 2603; RRID:AB_490795 Phospho-AMPKa (Thr172) Rabbit mAb Cell Signaling Cat#: 2535 Anti-rabbit IgG, HRP-linked Antibody Cell Signaling Cat#: 7074; RRID:AB_2099233 Anti-mouse IgG, HRP-linked Antibody Cell Signaling Cat#: 7076; RRID:AB_330924 Rabbit polyclonal anti-human/mouse beta-actin Cell Signaling Cat#: 4967; RRID:AB_330288 Chemicals, Peptides, and Recombinant Proteins IMDM GlutaMAX Thermo Fisher 3198-022 RPMI 1640 GlutaMAX Thermo Fisher 61870-010 X-Vivo 15 medium Lonza BE02-060F Seahorse XF DMEM medium, pH 7.4 Agilent 103575-100 Head-inactivated FCS Biochrom S 0115 – Lot# 1289W Penicillin-streptomycin Millipore P0781 2-mercaptoethanol Gibco #21985-023 Recombinant human TGF-beta Peprotech 100-21 (Continued on next page) Immunity 54, 1–16.e1–e6, January 12, 2021 e1 .. REAGENT or RESOURCE SOURCE IDENTIFIER Recombinant murine IL-6 Peprotech 211-11B Recombinant murine IL-1beta Peprotech 211-11B Recombinant murine IL-12 Peprotech 210-12 Recombinant murine IL-7 Peprotech 217-17 Recombinant murine IL-4 Preprotech 214-14 Recombinant human IL-12 p70 (HEK293 derived) Peprotech 200-12H Recombinant human IL-1beta R&D Systems 201-LB-005 Recombinant human IL-23 R&D Systems 1290-IL-010/CF Recombinant human IL-6 Peprotech 200-06 Recombinant human IL-21 Peprotech 200-21 Phorbol 12-myristate 13-acetate (PMA) Sigma-Aldrich P-8139 Ionomycin Sigma-Aldrich 10634 Tetramethzlyrhodamine, Ethyl Ester, Percholorate (TMRE) Invitrogen T669 Pierce RIPA Buffer Thermo Scientific 89900 Oligomycin Sigma Aldrich 75351 FCCP Cayman 15218 Rotenone Cayman 13955 Antimycin A Sigma Aldrich A8674 XF Base Medium Minimal DMEM without Phenol Red Agilent 103 Glucose R 99.5% D(+) Roth HN06.1 L-Glutamine Biochrom AG M11-004 Poly-D-lysine Hydrobromide Sigma Aldrich P6407 RNAse OUT Ribonuclease inhibitor (recombinant) Invitrogen #10777-019 CellTrace Violet Cell Proliferation Kit Life Technologies #65-0866-14 Fixable Viability Dye eFluor 780 eBioscience #65-0865-14 BD GolgiStop BD Biosciences #51-2092KZ FoxP3/transcription factor staining buffer set eBiosciences #00-5523-00 DMOG Cayman chemicals 71210 Lactate oxidase from Aerococcus viridans Sigma L9795-50UN Catalase, aspergillus niger Merck Millipore 219261-100KU Linezolid Chem-Impex 29723 Tigecycline hydrate Sigma PZ0021 Thiamphenicol Chem-Impex 29857 Vancomycin Cayman-Chemical #15327 FITC BrdU Flow Kit (RUO) BD 557891 DS18561882 inhibitor Medchemexpress HY-130251 Chaetocin from chaetomium minutum Merck C9492 DIF-1 Sigma SML0599 Mitomycin Cayman 11435 Paclitaxel Cayman 10461 SP600125 Inhibitor Cayman 10010466 ATP-polyamine-biotin Medchemexpress HY-D0183 Adenosine 5’triphosphate disodium salt hydrate Merck A26209 MCC905 InvivoGen inh-mcc (Continued on next page) e2 Immunity 54, 1–16.e1–e6, January 12, 2021
Functional Assay:Article Title: Metabolic Profiling Using Stable Isotope Tracing Reveals Distinct Patterns of Glucose Utilization by Physiologically Activated CD8 + T Cells.
Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Monoclonal anti-mouse CD3, eBioscience Functional Grade, Purified Invitrogen Clone # 145-2C11; RRID: AB_468849 Monoclonal anti-mouse CD28, eBioscience Functional Grade, Purified Invitrogen Clone # 37.51; RRID: AB_468923 Rat monoclonal anti-mouse CD25 eBioscience Clone # PC61.5 Rat monoclonal anti-mouse CD4 eBioscience Clone # GK1.5 Rat monoclonal anti-mouse CD62L eBioscience Clone # MEL-14 Rat monoclonal anti-mouse IFN-gamma eBioscience Clone # XMG1.2 Rat monoclonal anti-mouse CD8 eBioscience Clone # 53-6.7 Mouse monoclonal anti-mouse CD45.2 eBioscience Clone # 104 Rat monoclonal anti-mouse CD127 BioLegend Clone # A7R34; RRID AB_11218800 Hamster monoclonal anti-mouse KLRG1 eBioscience Clone # 2F1; RRID AB_1311265 Rat monoclonal anti-mouse CD44 eBioscience Clone # IM7 Rabbit polyclonal anti-human/mouse beta-actin Cell Signaling Cat # 4967; RRID AB_330288 Mouse monoclonal anti-mouse CD90.1 (Thy-1.1) eBioscience Clone # HIS51 Mouse monoclonal anti-mouse CD90.2 (Thy1-1.2) eBioscience Clone # 30-H12 Rabbit polyclonal anti mouse PHGDH Antibody ThermoFisher Scientific Cat # PA5-27578; RRID: AB_2545054 H-2Kb/OVA257 Tetramer Baylor College of Medicine N/A Bacterial and Virus Strains Listeria monocytogenes rLM-OVA van der Windt et al., 2012 N/A Luciferase shRNA (LMPd-Amt backbone): AGCTCCCGTGAATT GGAATCCTAGTGAAGCCACAGATGTAGGATTCCAATTCAGCG GGAGCC Chen et al., 2014 for vector backbone N/A Phgdh shRNA (LMPd-Amt backbone): tgctgttgacagtgagcgcgcact ggatgtgtttacagaatagtgaagccacagatgtattctgtaaacacatccagtgcatg cctactgcctcgga This paper N/A Chemicals, Peptides, and Recombinant Proteins [U-13C]-glucose Cambridge Isotopes Cat # CLM-1396 IMDM GlutaMAX Thermo Fisher Cat # 3198-022 Heat-inactivated FCS Biochrom Cat # 0115 – Lot# 1289W Penicillin-streptomycin Millipore Cat # A2213 2-mercaptoethanol GIBCO Cat # 21985-023 recombinant murine IL-2 Peprotech Cat # 212-12 recombinant murine IL-7 Peprotech Cat # 217-17 Phorbol 12-myristate 13-acetate (PMA) Sigma-Aldrich Cat # P-8139 Ionomycin Sigma-Aldrich Cat # 10634 FCCP Cayman Cat # 15218 Oligomycin Sigma-Aldrich Cat # 1404-19-9 Rotenone Cayman Cat # 13955 Antimycin A Sigma Aldrich Cat # A8674 XF Base Medium Minimal DMEM without Phenol Red Agilent Cat # 103335-100 Glucose R 99,5% D(+) Roth Cat # HN06.1 L-Glutamine Biochrom AG Cat # M11-004 Sodium Pyruvate GIBCO Cat # 11360-070 Poly-D-lysine hydrobromide Sigma Aldrich Cat # P6407 (Continued on next page) e1 Immunity 51, 1–15.e1–e5, November 19, 2019 ..
Purification:Article Title: Metabolic Profiling Using Stable Isotope Tracing Reveals Distinct Patterns of Glucose Utilization by Physiologically Activated CD8 + T Cells.
Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Monoclonal anti-mouse CD3, eBioscience Functional Grade, Purified Invitrogen Clone # 145-2C11; RRID: AB_468849 Monoclonal anti-mouse CD28, eBioscience Functional Grade, Purified Invitrogen Clone # 37.51; RRID: AB_468923 Rat monoclonal anti-mouse CD25 eBioscience Clone # PC61.5 Rat monoclonal anti-mouse CD4 eBioscience Clone # GK1.5 Rat monoclonal anti-mouse CD62L eBioscience Clone # MEL-14 Rat monoclonal anti-mouse IFN-gamma eBioscience Clone # XMG1.2 Rat monoclonal anti-mouse CD8 eBioscience Clone # 53-6.7 Mouse monoclonal anti-mouse CD45.2 eBioscience Clone # 104 Rat monoclonal anti-mouse CD127 BioLegend Clone # A7R34; RRID AB_11218800 Hamster monoclonal anti-mouse KLRG1 eBioscience Clone # 2F1; RRID AB_1311265 Rat monoclonal anti-mouse CD44 eBioscience Clone # IM7 Rabbit polyclonal anti-human/mouse beta-actin Cell Signaling Cat # 4967; RRID AB_330288 Mouse monoclonal anti-mouse CD90.1 (Thy-1.1) eBioscience Clone # HIS51 Mouse monoclonal anti-mouse CD90.2 (Thy1-1.2) eBioscience Clone # 30-H12 Rabbit polyclonal anti mouse PHGDH Antibody ThermoFisher Scientific Cat # PA5-27578; RRID: AB_2545054 H-2Kb/OVA257 Tetramer Baylor College of Medicine N/A Bacterial and Virus Strains Listeria monocytogenes rLM-OVA van der Windt et al., 2012 N/A Luciferase shRNA (LMPd-Amt backbone): AGCTCCCGTGAATT GGAATCCTAGTGAAGCCACAGATGTAGGATTCCAATTCAGCG GGAGCC Chen et al., 2014 for vector backbone N/A Phgdh shRNA (LMPd-Amt backbone): tgctgttgacagtgagcgcgcact ggatgtgtttacagaatagtgaagccacagatgtattctgtaaacacatccagtgcatg cctactgcctcgga This paper N/A Chemicals, Peptides, and Recombinant Proteins [U-13C]-glucose Cambridge Isotopes Cat # CLM-1396 IMDM GlutaMAX Thermo Fisher Cat # 3198-022 Heat-inactivated FCS Biochrom Cat # 0115 – Lot# 1289W Penicillin-streptomycin Millipore Cat # A2213 2-mercaptoethanol GIBCO Cat # 21985-023 recombinant murine IL-2 Peprotech Cat # 212-12 recombinant murine IL-7 Peprotech Cat # 217-17 Phorbol 12-myristate 13-acetate (PMA) Sigma-Aldrich Cat # P-8139 Ionomycin Sigma-Aldrich Cat # 10634 FCCP Cayman Cat # 15218 Oligomycin Sigma-Aldrich Cat # 1404-19-9 Rotenone Cayman Cat # 13955 Antimycin A Sigma Aldrich Cat # A8674 XF Base Medium Minimal DMEM without Phenol Red Agilent Cat # 103335-100 Glucose R 99,5% D(+) Roth Cat # HN06.1 L-Glutamine Biochrom AG Cat # M11-004 Sodium Pyruvate GIBCO Cat # 11360-070 Poly-D-lysine hydrobromide Sigma Aldrich Cat # P6407 (Continued on next page) e1 Immunity 51, 1–15.e1–e5, November 19, 2019 ..
Luciferase:Article Title: Metabolic Profiling Using Stable Isotope Tracing Reveals Distinct Patterns of Glucose Utilization by Physiologically Activated CD8 + T Cells.
Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Monoclonal anti-mouse CD3, eBioscience Functional Grade, Purified Invitrogen Clone # 145-2C11; RRID: AB_468849 Monoclonal anti-mouse CD28, eBioscience Functional Grade, Purified Invitrogen Clone # 37.51; RRID: AB_468923 Rat monoclonal anti-mouse CD25 eBioscience Clone # PC61.5 Rat monoclonal anti-mouse CD4 eBioscience Clone # GK1.5 Rat monoclonal anti-mouse CD62L eBioscience Clone # MEL-14 Rat monoclonal anti-mouse IFN-gamma eBioscience Clone # XMG1.2 Rat monoclonal anti-mouse CD8 eBioscience Clone # 53-6.7 Mouse monoclonal anti-mouse CD45.2 eBioscience Clone # 104 Rat monoclonal anti-mouse CD127 BioLegend Clone # A7R34; RRID AB_11218800 Hamster monoclonal anti-mouse KLRG1 eBioscience Clone # 2F1; RRID AB_1311265 Rat monoclonal anti-mouse CD44 eBioscience Clone # IM7 Rabbit polyclonal anti-human/mouse beta-actin Cell Signaling Cat # 4967; RRID AB_330288 Mouse monoclonal anti-mouse CD90.1 (Thy-1.1) eBioscience Clone # HIS51 Mouse monoclonal anti-mouse CD90.2 (Thy1-1.2) eBioscience Clone # 30-H12 Rabbit polyclonal anti mouse PHGDH Antibody ThermoFisher Scientific Cat # PA5-27578; RRID: AB_2545054 H-2Kb/OVA257 Tetramer Baylor College of Medicine N/A Bacterial and Virus Strains Listeria monocytogenes rLM-OVA van der Windt et al., 2012 N/A Luciferase shRNA (LMPd-Amt backbone): AGCTCCCGTGAATT GGAATCCTAGTGAAGCCACAGATGTAGGATTCCAATTCAGCG GGAGCC Chen et al., 2014 for vector backbone N/A Phgdh shRNA (LMPd-Amt backbone): tgctgttgacagtgagcgcgcact ggatgtgtttacagaatagtgaagccacagatgtattctgtaaacacatccagtgcatg cctactgcctcgga This paper N/A Chemicals, Peptides, and Recombinant Proteins [U-13C]-glucose Cambridge Isotopes Cat # CLM-1396 IMDM GlutaMAX Thermo Fisher Cat # 3198-022 Heat-inactivated FCS Biochrom Cat # 0115 – Lot# 1289W Penicillin-streptomycin Millipore Cat # A2213 2-mercaptoethanol GIBCO Cat # 21985-023 recombinant murine IL-2 Peprotech Cat # 212-12 recombinant murine IL-7 Peprotech Cat # 217-17 Phorbol 12-myristate 13-acetate (PMA) Sigma-Aldrich Cat # P-8139 Ionomycin Sigma-Aldrich Cat # 10634 FCCP Cayman Cat # 15218 Oligomycin Sigma-Aldrich Cat # 1404-19-9 Rotenone Cayman Cat # 13955 Antimycin A Sigma Aldrich Cat # A8674 XF Base Medium Minimal DMEM without Phenol Red Agilent Cat # 103335-100 Glucose R 99,5% D(+) Roth Cat # HN06.1 L-Glutamine Biochrom AG Cat # M11-004 Sodium Pyruvate GIBCO Cat # 11360-070 Poly-D-lysine hydrobromide Sigma Aldrich Cat # P6407 (Continued on next page) e1 Immunity 51, 1–15.e1–e5, November 19, 2019 ..
shRNA:Article Title: Metabolic Profiling Using Stable Isotope Tracing Reveals Distinct Patterns of Glucose Utilization by Physiologically Activated CD8 + T Cells.
Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Monoclonal anti-mouse CD3, eBioscience Functional Grade, Purified Invitrogen Clone # 145-2C11; RRID: AB_468849 Monoclonal anti-mouse CD28, eBioscience Functional Grade, Purified Invitrogen Clone # 37.51; RRID: AB_468923 Rat monoclonal anti-mouse CD25 eBioscience Clone # PC61.5 Rat monoclonal anti-mouse CD4 eBioscience Clone # GK1.5 Rat monoclonal anti-mouse CD62L eBioscience Clone # MEL-14 Rat monoclonal anti-mouse IFN-gamma eBioscience Clone # XMG1.2 Rat monoclonal anti-mouse CD8 eBioscience Clone # 53-6.7 Mouse monoclonal anti-mouse CD45.2 eBioscience Clone # 104 Rat monoclonal anti-mouse CD127 BioLegend Clone # A7R34; RRID AB_11218800 Hamster monoclonal anti-mouse KLRG1 eBioscience Clone # 2F1; RRID AB_1311265 Rat monoclonal anti-mouse CD44 eBioscience Clone # IM7 Rabbit polyclonal anti-human/mouse beta-actin Cell Signaling Cat # 4967; RRID AB_330288 Mouse monoclonal anti-mouse CD90.1 (Thy-1.1) eBioscience Clone # HIS51 Mouse monoclonal anti-mouse CD90.2 (Thy1-1.2) eBioscience Clone # 30-H12 Rabbit polyclonal anti mouse PHGDH Antibody ThermoFisher Scientific Cat # PA5-27578; RRID: AB_2545054 H-2Kb/OVA257 Tetramer Baylor College of Medicine N/A Bacterial and Virus Strains Listeria monocytogenes rLM-OVA van der Windt et al., 2012 N/A Luciferase shRNA (LMPd-Amt backbone): AGCTCCCGTGAATT GGAATCCTAGTGAAGCCACAGATGTAGGATTCCAATTCAGCG GGAGCC Chen et al., 2014 for vector backbone N/A Phgdh shRNA (LMPd-Amt backbone): tgctgttgacagtgagcgcgcact ggatgtgtttacagaatagtgaagccacagatgtattctgtaaacacatccagtgcatg cctactgcctcgga This paper N/A Chemicals, Peptides, and Recombinant Proteins [U-13C]-glucose Cambridge Isotopes Cat # CLM-1396 IMDM GlutaMAX Thermo Fisher Cat # 3198-022 Heat-inactivated FCS Biochrom Cat # 0115 – Lot# 1289W Penicillin-streptomycin Millipore Cat # A2213 2-mercaptoethanol GIBCO Cat # 21985-023 recombinant murine IL-2 Peprotech Cat # 212-12 recombinant murine IL-7 Peprotech Cat # 217-17 Phorbol 12-myristate 13-acetate (PMA) Sigma-Aldrich Cat # P-8139 Ionomycin Sigma-Aldrich Cat # 10634 FCCP Cayman Cat # 15218 Oligomycin Sigma-Aldrich Cat # 1404-19-9 Rotenone Cayman Cat # 13955 Antimycin A Sigma Aldrich Cat # A8674 XF Base Medium Minimal DMEM without Phenol Red Agilent Cat # 103335-100 Glucose R 99,5% D(+) Roth Cat # HN06.1 L-Glutamine Biochrom AG Cat # M11-004 Sodium Pyruvate GIBCO Cat # 11360-070 Poly-D-lysine hydrobromide Sigma Aldrich Cat # P6407 (Continued on next page) e1 Immunity 51, 1–15.e1–e5, November 19, 2019 ..
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