Western Blot:Article Title: Glutathione de novo synthesis but not recycling process coordinates with glutamine catabolism to control redox homeostasis and directs murine T cell differentiation
Article Snippet: Key resources table Reagent type (species) or resource Designation Source or reference Identifiers Additional information Strain, strain background (Mus musculus) C57BL/6 (B6) mice Taconic Strain, strain background (Mus musculus) C3H/HeN Envigo Continued on next page Lian et al. eLife 2018;7:e36158. .. DOI: https://doi.org/10.7554/eLife.36158 16 of 28 Continued Reagent type (species) or resource Designation Source or reference Identifiers Additional information Genetic reagent (Mus musculus) CD4-Cre Gclcflox/flox PMID:23226398 Genetic reagent (Mus musculus) Gclm-KO PMID:12384496 Genetic reagent (Mus musculus) ROSA26CreERT2 RRID: IMSR_JAX:008463 The Jackson Laboratory Genetic reagent (Mus musculus) GSR-KO PMID: 10218442 Antibody Mouse antiCD3 mAb Cat. #:BE0001-1, RRID:AB_1107634 BioXcell Antibody Mouse antiCD28 mAb Cat. #BE0015-1 RRID:AB_1107624 BioXcell Antibody Mouse anti -IL2 mAb Cat.#BE0043 RRID::AB_1107702 BioXcell Antibody Mouse antiIL4 mAb Cat. #BE0045 RRID:AB_1107707 BioXcell Antibody Mouse antiIFNg mAb Cat. #BE0055 RRID:AB_1107694 BioXcell Antibody Anti mouse CD4-FITC Cat. #11–0042 RRID:AB_464897 eBioscience (1:200) Antibody Anti mouse CD4-APC Cat. #17-0041-81 RRID:AB_469319 eBioscience (1:200) Antibody Anti mouse CD8-APC-Cy7 Cat. #100714 RRID:AB_312753 Biolegend (1:200) Antibody Anti mouse Foxp3-APC Cat. # RRID:AB_469456 eBioscience (1:200) Antibody Anti mouse IL-17A-PECy7 Cat. #25-7177-82 RRID:AB_10732356 eBioscience (1:200) Antibody Anti GCLC antibody (rabbit monoclonal) Cat. #ab190685 RRID:AB_10975474 Abcam WB (1:1000) Antibody Anti GCLM antibody (rabbit monoclonal) Cat. #ab124827 RRID:AB_10975474 Abcam WB (1:1000) Antibody anti-mouse CD25 -PE Cat. #101904 RRID:AB_312847 Biolegend (1:200) Antibody anti-mouse CD69-PECy7 Cat. #552879 RRID:AB_394508 BD Bioscience (1:200) Antibody Anti mouse monoclonal CD3 Cat. #sc-101442 RRID:AB_1120355 Santa Cruz IHC (1:50) Antibody Anti mouse monoclonal galectin-3 Cat. #sc-32790, RRID:AB_627657 Santa Cruz IHC (1:50) Peptide, recombinant protein MOG35-55 peptide synthesized and HPLC-purified St. Jude Hartwell Center for Biotechnology Peptide, recombinant protein Recombinant mouse IL-6 216–16 Peprotech Peptide, recombinant protein Recombinant human TGFb 100–21 c Peprotech Continued on next page Lian et al. eLife 2018;7:e36158. .. DOI: https://doi.org/10.7554/eLife.36158 17 of 28 Continued Reagent type (species) or resource Designation Source or reference Identifiers Additional information Peptide, recombinant protein Recombinant human or mouse IL-2 200–02 Peprotech Commercial assay or kit Foxp3/Transcription Factor Staining Buffer Set 00-5523-00 e-Bioscience Commercial assay or kit Naive CD4 + T cell isolation kit,mouse 5160725186 Miltenyi Biotec Commercial assay or kit CD45R(B220) microbeads, mouse 5150309030 Miltenyi Biotec Commercial assay or kit ABC kit PK-7200 Vector laboratories Commercial assay or kit MojoSort Mouse naive CD4 T Cell Isolation Kit 480031 Biolegend Chemical compound, drug Diethly Fumerate Sigma Aldrich D95654 Chemical compound, drug N-Acetyl-L-cysteine Sigma-Aldrich A7250 Chemical compound, drug Tamofixen Sigma-Aldrich T5648 Chemical compound, drug 4-hydroxytamoxifen Sigma H7904 Chemical compound, drug Dimethy a-keto glutarate/aKG Sigma-Aldrich 34963–1 Chemical compound, drug Hypoxathine Sigma-Aldrich H9377 Chemical compound, drug Thymidine Sigma T9250 Chemical compound, drug H2O2 Sigma-Aldrich 7722-84-1 Chemical compound, drug carboxyfluorescein diacetate succinimidyl ester(CFSE) Invitrogen C1157 Chemical compound, drug DM-H2DCFDA Invitrogen C6827 Chemical compound, drug DAB Vector Laboratories SK-4100 Chemical compound, drug Monobromobimane Invitrogen M1378 Chemical compound, drug 7-aminoactinomycin D(7AAD) Biolegend 420404 Chemical compound, drug Pertussis toxin 181 List Biological Laboratories Chemical compound, drug Mycobacterium tuberculosum 231141 Difco Chemical compound, drug Incomplete Freund’s Adjuvant 263910 Difco Chemical compound, drug [U-14C]-glutamine MC 1124 Moravek Chemical compound, drug [2–14C]-pyruvate ARC 0222 American Radiolabeled Chemicals Continued on next page Lian et al. eLife 2018;7:e36158.
Immunohistochemistry:Article Title: Glutathione de novo synthesis but not recycling process coordinates with glutamine catabolism to control redox homeostasis and directs murine T cell differentiation
Article Snippet: Key resources table Reagent type (species) or resource Designation Source or reference Identifiers Additional information Strain, strain background (Mus musculus) C57BL/6 (B6) mice Taconic Strain, strain background (Mus musculus) C3H/HeN Envigo Continued on next page Lian et al. eLife 2018;7:e36158. .. DOI: https://doi.org/10.7554/eLife.36158 16 of 28 Continued Reagent type (species) or resource Designation Source or reference Identifiers Additional information Genetic reagent (Mus musculus) CD4-Cre Gclcflox/flox PMID:23226398 Genetic reagent (Mus musculus) Gclm-KO PMID:12384496 Genetic reagent (Mus musculus) ROSA26CreERT2 RRID: IMSR_JAX:008463 The Jackson Laboratory Genetic reagent (Mus musculus) GSR-KO PMID: 10218442 Antibody Mouse antiCD3 mAb Cat. #:BE0001-1, RRID:AB_1107634 BioXcell Antibody Mouse antiCD28 mAb Cat. #BE0015-1 RRID:AB_1107624 BioXcell Antibody Mouse anti -IL2 mAb Cat.#BE0043 RRID::AB_1107702 BioXcell Antibody Mouse antiIL4 mAb Cat. #BE0045 RRID:AB_1107707 BioXcell Antibody Mouse antiIFNg mAb Cat. #BE0055 RRID:AB_1107694 BioXcell Antibody Anti mouse CD4-FITC Cat. #11–0042 RRID:AB_464897 eBioscience (1:200) Antibody Anti mouse CD4-APC Cat. #17-0041-81 RRID:AB_469319 eBioscience (1:200) Antibody Anti mouse CD8-APC-Cy7 Cat. #100714 RRID:AB_312753 Biolegend (1:200) Antibody Anti mouse Foxp3-APC Cat. # RRID:AB_469456 eBioscience (1:200) Antibody Anti mouse IL-17A-PECy7 Cat. #25-7177-82 RRID:AB_10732356 eBioscience (1:200) Antibody Anti GCLC antibody (rabbit monoclonal) Cat. #ab190685 RRID:AB_10975474 Abcam WB (1:1000) Antibody Anti GCLM antibody (rabbit monoclonal) Cat. #ab124827 RRID:AB_10975474 Abcam WB (1:1000) Antibody anti-mouse CD25 -PE Cat. #101904 RRID:AB_312847 Biolegend (1:200) Antibody anti-mouse CD69-PECy7 Cat. #552879 RRID:AB_394508 BD Bioscience (1:200) Antibody Anti mouse monoclonal CD3 Cat. #sc-101442 RRID:AB_1120355 Santa Cruz IHC (1:50) Antibody Anti mouse monoclonal galectin-3 Cat. #sc-32790, RRID:AB_627657 Santa Cruz IHC (1:50) Peptide, recombinant protein MOG35-55 peptide synthesized and HPLC-purified St. Jude Hartwell Center for Biotechnology Peptide, recombinant protein Recombinant mouse IL-6 216–16 Peprotech Peptide, recombinant protein Recombinant human TGFb 100–21 c Peprotech Continued on next page Lian et al. eLife 2018;7:e36158. .. DOI: https://doi.org/10.7554/eLife.36158 17 of 28 Continued Reagent type (species) or resource Designation Source or reference Identifiers Additional information Peptide, recombinant protein Recombinant human or mouse IL-2 200–02 Peprotech Commercial assay or kit Foxp3/Transcription Factor Staining Buffer Set 00-5523-00 e-Bioscience Commercial assay or kit Naive CD4 + T cell isolation kit,mouse 5160725186 Miltenyi Biotec Commercial assay or kit CD45R(B220) microbeads, mouse 5150309030 Miltenyi Biotec Commercial assay or kit ABC kit PK-7200 Vector laboratories Commercial assay or kit MojoSort Mouse naive CD4 T Cell Isolation Kit 480031 Biolegend Chemical compound, drug Diethly Fumerate Sigma Aldrich D95654 Chemical compound, drug N-Acetyl-L-cysteine Sigma-Aldrich A7250 Chemical compound, drug Tamofixen Sigma-Aldrich T5648 Chemical compound, drug 4-hydroxytamoxifen Sigma H7904 Chemical compound, drug Dimethy a-keto glutarate/aKG Sigma-Aldrich 34963–1 Chemical compound, drug Hypoxathine Sigma-Aldrich H9377 Chemical compound, drug Thymidine Sigma T9250 Chemical compound, drug H2O2 Sigma-Aldrich 7722-84-1 Chemical compound, drug carboxyfluorescein diacetate succinimidyl ester(CFSE) Invitrogen C1157 Chemical compound, drug DM-H2DCFDA Invitrogen C6827 Chemical compound, drug DAB Vector Laboratories SK-4100 Chemical compound, drug Monobromobimane Invitrogen M1378 Chemical compound, drug 7-aminoactinomycin D(7AAD) Biolegend 420404 Chemical compound, drug Pertussis toxin 181 List Biological Laboratories Chemical compound, drug Mycobacterium tuberculosum 231141 Difco Chemical compound, drug Incomplete Freund’s Adjuvant 263910 Difco Chemical compound, drug [U-14C]-glutamine MC 1124 Moravek Chemical compound, drug [2–14C]-pyruvate ARC 0222 American Radiolabeled Chemicals Continued on next page Lian et al. eLife 2018;7:e36158.
Recombinant:Article Title: Glutathione de novo synthesis but not recycling process coordinates with glutamine catabolism to control redox homeostasis and directs murine T cell differentiation
Article Snippet: Key resources table Reagent type (species) or resource Designation Source or reference Identifiers Additional information Strain, strain background (Mus musculus) C57BL/6 (B6) mice Taconic Strain, strain background (Mus musculus) C3H/HeN Envigo Continued on next page Lian et al. eLife 2018;7:e36158. .. DOI: https://doi.org/10.7554/eLife.36158 16 of 28 Continued Reagent type (species) or resource Designation Source or reference Identifiers Additional information Genetic reagent (Mus musculus) CD4-Cre Gclcflox/flox PMID:23226398 Genetic reagent (Mus musculus) Gclm-KO PMID:12384496 Genetic reagent (Mus musculus) ROSA26CreERT2 RRID: IMSR_JAX:008463 The Jackson Laboratory Genetic reagent (Mus musculus) GSR-KO PMID: 10218442 Antibody Mouse antiCD3 mAb Cat. #:BE0001-1, RRID:AB_1107634 BioXcell Antibody Mouse antiCD28 mAb Cat. #BE0015-1 RRID:AB_1107624 BioXcell Antibody Mouse anti -IL2 mAb Cat.#BE0043 RRID::AB_1107702 BioXcell Antibody Mouse antiIL4 mAb Cat. #BE0045 RRID:AB_1107707 BioXcell Antibody Mouse antiIFNg mAb Cat. #BE0055 RRID:AB_1107694 BioXcell Antibody Anti mouse CD4-FITC Cat. #11–0042 RRID:AB_464897 eBioscience (1:200) Antibody Anti mouse CD4-APC Cat. #17-0041-81 RRID:AB_469319 eBioscience (1:200) Antibody Anti mouse CD8-APC-Cy7 Cat. #100714 RRID:AB_312753 Biolegend (1:200) Antibody Anti mouse Foxp3-APC Cat. # RRID:AB_469456 eBioscience (1:200) Antibody Anti mouse IL-17A-PECy7 Cat. #25-7177-82 RRID:AB_10732356 eBioscience (1:200) Antibody Anti GCLC antibody (rabbit monoclonal) Cat. #ab190685 RRID:AB_10975474 Abcam WB (1:1000) Antibody Anti GCLM antibody (rabbit monoclonal) Cat. #ab124827 RRID:AB_10975474 Abcam WB (1:1000) Antibody anti-mouse CD25 -PE Cat. #101904 RRID:AB_312847 Biolegend (1:200) Antibody anti-mouse CD69-PECy7 Cat. #552879 RRID:AB_394508 BD Bioscience (1:200) Antibody Anti mouse monoclonal CD3 Cat. #sc-101442 RRID:AB_1120355 Santa Cruz IHC (1:50) Antibody Anti mouse monoclonal galectin-3 Cat. #sc-32790, RRID:AB_627657 Santa Cruz IHC (1:50) Peptide, recombinant protein MOG35-55 peptide synthesized and HPLC-purified St. Jude Hartwell Center for Biotechnology Peptide, recombinant protein Recombinant mouse IL-6 216–16 Peprotech Peptide, recombinant protein Recombinant human TGFb 100–21 c Peprotech Continued on next page Lian et al. eLife 2018;7:e36158. .. DOI: https://doi.org/10.7554/eLife.36158 17 of 28 Continued Reagent type (species) or resource Designation Source or reference Identifiers Additional information Peptide, recombinant protein Recombinant human or mouse IL-2 200–02 Peprotech Commercial assay or kit Foxp3/Transcription Factor Staining Buffer Set 00-5523-00 e-Bioscience Commercial assay or kit Naive CD4 + T cell isolation kit,mouse 5160725186 Miltenyi Biotec Commercial assay or kit CD45R(B220) microbeads, mouse 5150309030 Miltenyi Biotec Commercial assay or kit ABC kit PK-7200 Vector laboratories Commercial assay or kit MojoSort Mouse naive CD4 T Cell Isolation Kit 480031 Biolegend Chemical compound, drug Diethly Fumerate Sigma Aldrich D95654 Chemical compound, drug N-Acetyl-L-cysteine Sigma-Aldrich A7250 Chemical compound, drug Tamofixen Sigma-Aldrich T5648 Chemical compound, drug 4-hydroxytamoxifen Sigma H7904 Chemical compound, drug Dimethy a-keto glutarate/aKG Sigma-Aldrich 34963–1 Chemical compound, drug Hypoxathine Sigma-Aldrich H9377 Chemical compound, drug Thymidine Sigma T9250 Chemical compound, drug H2O2 Sigma-Aldrich 7722-84-1 Chemical compound, drug carboxyfluorescein diacetate succinimidyl ester(CFSE) Invitrogen C1157 Chemical compound, drug DM-H2DCFDA Invitrogen C6827 Chemical compound, drug DAB Vector Laboratories SK-4100 Chemical compound, drug Monobromobimane Invitrogen M1378 Chemical compound, drug 7-aminoactinomycin D(7AAD) Biolegend 420404 Chemical compound, drug Pertussis toxin 181 List Biological Laboratories Chemical compound, drug Mycobacterium tuberculosum 231141 Difco Chemical compound, drug Incomplete Freund’s Adjuvant 263910 Difco Chemical compound, drug [U-14C]-glutamine MC 1124 Moravek Chemical compound, drug [2–14C]-pyruvate ARC 0222 American Radiolabeled Chemicals Continued on next page Lian et al. eLife 2018;7:e36158.
Synthesized:Article Title: Glutathione de novo synthesis but not recycling process coordinates with glutamine catabolism to control redox homeostasis and directs murine T cell differentiation
Article Snippet: Key resources table Reagent type (species) or resource Designation Source or reference Identifiers Additional information Strain, strain background (Mus musculus) C57BL/6 (B6) mice Taconic Strain, strain background (Mus musculus) C3H/HeN Envigo Continued on next page Lian et al. eLife 2018;7:e36158. .. DOI: https://doi.org/10.7554/eLife.36158 16 of 28 Continued Reagent type (species) or resource Designation Source or reference Identifiers Additional information Genetic reagent (Mus musculus) CD4-Cre Gclcflox/flox PMID:23226398 Genetic reagent (Mus musculus) Gclm-KO PMID:12384496 Genetic reagent (Mus musculus) ROSA26CreERT2 RRID: IMSR_JAX:008463 The Jackson Laboratory Genetic reagent (Mus musculus) GSR-KO PMID: 10218442 Antibody Mouse antiCD3 mAb Cat. #:BE0001-1, RRID:AB_1107634 BioXcell Antibody Mouse antiCD28 mAb Cat. #BE0015-1 RRID:AB_1107624 BioXcell Antibody Mouse anti -IL2 mAb Cat.#BE0043 RRID::AB_1107702 BioXcell Antibody Mouse antiIL4 mAb Cat. #BE0045 RRID:AB_1107707 BioXcell Antibody Mouse antiIFNg mAb Cat. #BE0055 RRID:AB_1107694 BioXcell Antibody Anti mouse CD4-FITC Cat. #11–0042 RRID:AB_464897 eBioscience (1:200) Antibody Anti mouse CD4-APC Cat. #17-0041-81 RRID:AB_469319 eBioscience (1:200) Antibody Anti mouse CD8-APC-Cy7 Cat. #100714 RRID:AB_312753 Biolegend (1:200) Antibody Anti mouse Foxp3-APC Cat. # RRID:AB_469456 eBioscience (1:200) Antibody Anti mouse IL-17A-PECy7 Cat. #25-7177-82 RRID:AB_10732356 eBioscience (1:200) Antibody Anti GCLC antibody (rabbit monoclonal) Cat. #ab190685 RRID:AB_10975474 Abcam WB (1:1000) Antibody Anti GCLM antibody (rabbit monoclonal) Cat. #ab124827 RRID:AB_10975474 Abcam WB (1:1000) Antibody anti-mouse CD25 -PE Cat. #101904 RRID:AB_312847 Biolegend (1:200) Antibody anti-mouse CD69-PECy7 Cat. #552879 RRID:AB_394508 BD Bioscience (1:200) Antibody Anti mouse monoclonal CD3 Cat. #sc-101442 RRID:AB_1120355 Santa Cruz IHC (1:50) Antibody Anti mouse monoclonal galectin-3 Cat. #sc-32790, RRID:AB_627657 Santa Cruz IHC (1:50) Peptide, recombinant protein MOG35-55 peptide synthesized and HPLC-purified St. Jude Hartwell Center for Biotechnology Peptide, recombinant protein Recombinant mouse IL-6 216–16 Peprotech Peptide, recombinant protein Recombinant human TGFb 100–21 c Peprotech Continued on next page Lian et al. eLife 2018;7:e36158. .. DOI: https://doi.org/10.7554/eLife.36158 17 of 28 Continued Reagent type (species) or resource Designation Source or reference Identifiers Additional information Peptide, recombinant protein Recombinant human or mouse IL-2 200–02 Peprotech Commercial assay or kit Foxp3/Transcription Factor Staining Buffer Set 00-5523-00 e-Bioscience Commercial assay or kit Naive CD4 + T cell isolation kit,mouse 5160725186 Miltenyi Biotec Commercial assay or kit CD45R(B220) microbeads, mouse 5150309030 Miltenyi Biotec Commercial assay or kit ABC kit PK-7200 Vector laboratories Commercial assay or kit MojoSort Mouse naive CD4 T Cell Isolation Kit 480031 Biolegend Chemical compound, drug Diethly Fumerate Sigma Aldrich D95654 Chemical compound, drug N-Acetyl-L-cysteine Sigma-Aldrich A7250 Chemical compound, drug Tamofixen Sigma-Aldrich T5648 Chemical compound, drug 4-hydroxytamoxifen Sigma H7904 Chemical compound, drug Dimethy a-keto glutarate/aKG Sigma-Aldrich 34963–1 Chemical compound, drug Hypoxathine Sigma-Aldrich H9377 Chemical compound, drug Thymidine Sigma T9250 Chemical compound, drug H2O2 Sigma-Aldrich 7722-84-1 Chemical compound, drug carboxyfluorescein diacetate succinimidyl ester(CFSE) Invitrogen C1157 Chemical compound, drug DM-H2DCFDA Invitrogen C6827 Chemical compound, drug DAB Vector Laboratories SK-4100 Chemical compound, drug Monobromobimane Invitrogen M1378 Chemical compound, drug 7-aminoactinomycin D(7AAD) Biolegend 420404 Chemical compound, drug Pertussis toxin 181 List Biological Laboratories Chemical compound, drug Mycobacterium tuberculosum 231141 Difco Chemical compound, drug Incomplete Freund’s Adjuvant 263910 Difco Chemical compound, drug [U-14C]-glutamine MC 1124 Moravek Chemical compound, drug [2–14C]-pyruvate ARC 0222 American Radiolabeled Chemicals Continued on next page Lian et al. eLife 2018;7:e36158.
High Performance Liquid Chromatography:Article Title: Glutathione de novo synthesis but not recycling process coordinates with glutamine catabolism to control redox homeostasis and directs murine T cell differentiation
Article Snippet: Key resources table Reagent type (species) or resource Designation Source or reference Identifiers Additional information Strain, strain background (Mus musculus) C57BL/6 (B6) mice Taconic Strain, strain background (Mus musculus) C3H/HeN Envigo Continued on next page Lian et al. eLife 2018;7:e36158. .. DOI: https://doi.org/10.7554/eLife.36158 16 of 28 Continued Reagent type (species) or resource Designation Source or reference Identifiers Additional information Genetic reagent (Mus musculus) CD4-Cre Gclcflox/flox PMID:23226398 Genetic reagent (Mus musculus) Gclm-KO PMID:12384496 Genetic reagent (Mus musculus) ROSA26CreERT2 RRID: IMSR_JAX:008463 The Jackson Laboratory Genetic reagent (Mus musculus) GSR-KO PMID: 10218442 Antibody Mouse antiCD3 mAb Cat. #:BE0001-1, RRID:AB_1107634 BioXcell Antibody Mouse antiCD28 mAb Cat. #BE0015-1 RRID:AB_1107624 BioXcell Antibody Mouse anti -IL2 mAb Cat.#BE0043 RRID::AB_1107702 BioXcell Antibody Mouse antiIL4 mAb Cat. #BE0045 RRID:AB_1107707 BioXcell Antibody Mouse antiIFNg mAb Cat. #BE0055 RRID:AB_1107694 BioXcell Antibody Anti mouse CD4-FITC Cat. #11–0042 RRID:AB_464897 eBioscience (1:200) Antibody Anti mouse CD4-APC Cat. #17-0041-81 RRID:AB_469319 eBioscience (1:200) Antibody Anti mouse CD8-APC-Cy7 Cat. #100714 RRID:AB_312753 Biolegend (1:200) Antibody Anti mouse Foxp3-APC Cat. # RRID:AB_469456 eBioscience (1:200) Antibody Anti mouse IL-17A-PECy7 Cat. #25-7177-82 RRID:AB_10732356 eBioscience (1:200) Antibody Anti GCLC antibody (rabbit monoclonal) Cat. #ab190685 RRID:AB_10975474 Abcam WB (1:1000) Antibody Anti GCLM antibody (rabbit monoclonal) Cat. #ab124827 RRID:AB_10975474 Abcam WB (1:1000) Antibody anti-mouse CD25 -PE Cat. #101904 RRID:AB_312847 Biolegend (1:200) Antibody anti-mouse CD69-PECy7 Cat. #552879 RRID:AB_394508 BD Bioscience (1:200) Antibody Anti mouse monoclonal CD3 Cat. #sc-101442 RRID:AB_1120355 Santa Cruz IHC (1:50) Antibody Anti mouse monoclonal galectin-3 Cat. #sc-32790, RRID:AB_627657 Santa Cruz IHC (1:50) Peptide, recombinant protein MOG35-55 peptide synthesized and HPLC-purified St. Jude Hartwell Center for Biotechnology Peptide, recombinant protein Recombinant mouse IL-6 216–16 Peprotech Peptide, recombinant protein Recombinant human TGFb 100–21 c Peprotech Continued on next page Lian et al. eLife 2018;7:e36158. .. DOI: https://doi.org/10.7554/eLife.36158 17 of 28 Continued Reagent type (species) or resource Designation Source or reference Identifiers Additional information Peptide, recombinant protein Recombinant human or mouse IL-2 200–02 Peprotech Commercial assay or kit Foxp3/Transcription Factor Staining Buffer Set 00-5523-00 e-Bioscience Commercial assay or kit Naive CD4 + T cell isolation kit,mouse 5160725186 Miltenyi Biotec Commercial assay or kit CD45R(B220) microbeads, mouse 5150309030 Miltenyi Biotec Commercial assay or kit ABC kit PK-7200 Vector laboratories Commercial assay or kit MojoSort Mouse naive CD4 T Cell Isolation Kit 480031 Biolegend Chemical compound, drug Diethly Fumerate Sigma Aldrich D95654 Chemical compound, drug N-Acetyl-L-cysteine Sigma-Aldrich A7250 Chemical compound, drug Tamofixen Sigma-Aldrich T5648 Chemical compound, drug 4-hydroxytamoxifen Sigma H7904 Chemical compound, drug Dimethy a-keto glutarate/aKG Sigma-Aldrich 34963–1 Chemical compound, drug Hypoxathine Sigma-Aldrich H9377 Chemical compound, drug Thymidine Sigma T9250 Chemical compound, drug H2O2 Sigma-Aldrich 7722-84-1 Chemical compound, drug carboxyfluorescein diacetate succinimidyl ester(CFSE) Invitrogen C1157 Chemical compound, drug DM-H2DCFDA Invitrogen C6827 Chemical compound, drug DAB Vector Laboratories SK-4100 Chemical compound, drug Monobromobimane Invitrogen M1378 Chemical compound, drug 7-aminoactinomycin D(7AAD) Biolegend 420404 Chemical compound, drug Pertussis toxin 181 List Biological Laboratories Chemical compound, drug Mycobacterium tuberculosum 231141 Difco Chemical compound, drug Incomplete Freund’s Adjuvant 263910 Difco Chemical compound, drug [U-14C]-glutamine MC 1124 Moravek Chemical compound, drug [2–14C]-pyruvate ARC 0222 American Radiolabeled Chemicals Continued on next page Lian et al. eLife 2018;7:e36158.
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