Enzyme-linked Immunosorbent Assay:Article Title: JAK1 signaling in dendritic cells promotes peripheral tolerance in autoimmunity through PD-L1-mediated regulatory T cell induction.
Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Chemicals, peptides, and recombinant proteins Myelin oligodendrocyte glycoprotein (MOG)35–55 Charitè Berlin NA Incomplete Freund’s adjuvant Sigma-Aldrich Cat# F5506 Mycobacterium tuberculosis H37Ra BD Cat# 231141 Pertussis toxin List Biological Laboratories Cat# 181 recombinant murine GM-CSF R&D Cat# 415-ML-050 recombinant murine IL-4 Miltenyi Biotec Cat# 130-097-761 recombinant murine IFN-ɣ Miltenyi Biotec Cat# 130-105-774 recombinant murine IL-10 R&D Cat# 417-ML-005 Lipopolysaccharide ultra pure InvivoGen Cat# tlrl-pelps Filgotinib TargetMol Cat# T1929 Stattic MedChem Express Cat# HY-13818 Fludarabine MedChem Express Cat# HY-B0069 Ruxolitinib Cayman Chemical Cat# 11609-5 Tofacitinib MedChem Express Cat# HY-40354 OVA323–399 Bachem Cat# H-5532 CD11c MACS Micro Bead System Miltenyi Biotec Cat# 130-125-835 Naive CD4+ T Cell Isolation Kit, mouse Miltenyi Biotec Cat# 130-104-453 CFSE Thermo Fisher Scientific Cat# 65-0850-84 Recombinant Mouse TGF-beta 1 R&D Cat# 7666-MB-005/CF Animal-Free Recombinant Human IL-2 Peprotech Cat# AF-200-02 Recombinant mouse PD-L1 Fc-Fusion protein Kerafast Cat# Pr00112-1.9 Recombinant human IgG1 Fc control protein R&D Cat# 110-HG-100 Adult brain dissociation kit Miltenyi Biotec Cat# 130-107-677 Fixable Viability Dye eFluorTM 780 Thermo Fisher Scientific Cat# 65-0865-14 SYTOXTM AADvancedTM Thermo Fisher Scientific Cat# S10274 FoxP3/Transcription factor staining buffer set Thermo Fisher Scientific Cat# 00-5523-00 eBioscienceTM Intracellular fixation & permeabilization buffer set Thermo Fisher Scientific Cat# 88-8824-00 True-Phos Buffer Set BioLegend Cat# 425401 Ionomycin Sigma Aldrich Cat# I0634 PMA Sigma Aldrich Cat# P1585 Brefeldin A solution Thermo Fisher Scientific Cat# 00-4506-51 High-Capacity cDNA Reverse Transcription Kit Thermo Fisher Scientific Cat# 4368814 TaqManTM Gene Expression Master Mix Thermo Fisher Scientific Cat# 4369016 Luna Master Mix New England BioLabs Cat# M3003E Critical commercial assays LEGENDplexTM Mouse Inflammation Panel BioLegend Cat# 740446 LEGENDplexTM Mouse Th Panel BioLegend Cat# 740741 Experimental models: Organisms/strains Mouse: Jak1tm1a(EUCOMM)Hmgu EUMMCR MGI:4456720 Mouse: ROSA26::FLPe knock in; B6.129S4-Gt(ROSA)26Sortm1(FLP1)Dym/RainJ The Jackson Laboratory Cat# JAX:009086, RRID:IMSR_JAX:009086 Mouse: CD11c cre; B6.Cg-Tg(Itgax-cre)11Reiz/J mice The Jackson Laboratory Cat# JAX:008068, RRID:IMSR_JAX:008068 (Continued on next page) Cell Reports 38, 110420, February 22, 2022 e3
Article Title: Succinate uptake by T cells suppresses their effector function via inhibition of mitochondrial glucose oxidation.
Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies anti-CD69-APC Biolegend Clone FN50, Cat# 310910; RRID:AB_314845 anti-CD25-BV605 Biolegend Clone BC96, Cat# 302632; RRID:AB_11218989 anti-CD107a-FITC BD Pharmingen Clone H4A3, Cat# 560949; RRID:AB_2033934 anti-IFN-g-FITC Biolegend Clone B27, Cat# 506504; RRID:AB_315437 anti-TNF-a-PE Biolegend Clone Mab11, Cat# 502909; RRID:AB_315261 anti-IFN-g capture (ELISA) Bio-Rad Clone AbD00676, Cat# HCA043; RRID:AB_906045 anti-IFN-g biotinylated (ELISA) Bio-Rad Clone 2503 Cat# HCA044P; RRID:AB_10614712 anti-HIF-1a [H1alpha67] (WB) Abcam Cat# ab1; RRID:AB_296474 anti-SUCNR1 (WB) Novus Biologicals Cat# NBP1-00861; RRID:AB_1503315 anti-CD3 (mouse) Biolegend Clone 145-2C11 Cat# 100340; RRID:AB_11149115 anti-CD28 (mouse) Biolegend Clone 37.51 Cat# 102116; RRID:AB_11147170 anti-IFN-g-FITC (mouse) Biolegend Clone XMG1.2, Cat# 505806; RRID:AB_315400 anti-TNF-a-PE (mouse) Biolegend Clone MP6-XT22, Cat# 506305; RRID:AB_315426 Chemicals, peptides, and recombinant proteins RPMI1640 Cell Culture Medium Sigma Aldrich Cat# F9665 Penicillin/Streptomycin Thermo Fisher Cat# 15140122 Recombinant human IL-2 PeproTech Cat# 200-02 Immunocult Human CD3/CD28 T cell activator Stemcell Technologies Cat# 10991 Sodium Succinate Sigma Aldrich Cat# 14160 AZD-3965 Selleckchem Cat# S7339 b-chloro-L-alanine Sigma Aldrich Cat# C9033 Sodium Pyruvate Sigma Aldrich Cat# P8574 Dichloroacetate Sigma Aldrich Cat# 347795 Sodium Metavanadate Sigma Aldrich Cat# 72060 Cell Activation Cocktail Biolegend Cat# 423304 Fixation/permeabilization solutions eBioscience Cat# 00-5523-00 CellTrace Violet Thermo Fisher Cat# C34557 Recombinant IFN-g standard (ELISA) Bio-Rad Cat# PHP050 streptavidin-HRP Sigma Aldrich Cat# E2866 TMB substrate Bio-Rad Cat# BUF062B MitoSpy Orange Biolegend Cat# 424804 Laemmli buffer Sigma Aldrich Cat# 38733 RIPA buffer Thermo Fisher Scientific Cat# 89900 Clarity Western ECL Substrate Bio-Rad Cat# 170-5061 U-13C6 glucose CK Isotopes Cat# CLM-1396-1 13C4 sodium succinate Sigma Aldrich Cat# 491985 Methoxyamine hydrochloride in pyridine Thermo Fisher Scientific Cat# 25104 Lipofectamine RNAiMAX Thermo Fisher Cat# 13778075 Fixable Viability Dye eFluorTM 780 Thermo Fisher Scientific Cat# 65-0865-14 4C (SUCNR1 antagonist) Provided by Prof S Pluchino N/A (Continued on next page) Cell Reports 40, 111193, August 16, 2022 e1
Western Blot:Article Title: JAK1 signaling in dendritic cells promotes peripheral tolerance in autoimmunity through PD-L1-mediated regulatory T cell induction.
Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Chemicals, peptides, and recombinant proteins Myelin oligodendrocyte glycoprotein (MOG)35–55 Charitè Berlin NA Incomplete Freund’s adjuvant Sigma-Aldrich Cat# F5506 Mycobacterium tuberculosis H37Ra BD Cat# 231141 Pertussis toxin List Biological Laboratories Cat# 181 recombinant murine GM-CSF R&D Cat# 415-ML-050 recombinant murine IL-4 Miltenyi Biotec Cat# 130-097-761 recombinant murine IFN-ɣ Miltenyi Biotec Cat# 130-105-774 recombinant murine IL-10 R&D Cat# 417-ML-005 Lipopolysaccharide ultra pure InvivoGen Cat# tlrl-pelps Filgotinib TargetMol Cat# T1929 Stattic MedChem Express Cat# HY-13818 Fludarabine MedChem Express Cat# HY-B0069 Ruxolitinib Cayman Chemical Cat# 11609-5 Tofacitinib MedChem Express Cat# HY-40354 OVA323–399 Bachem Cat# H-5532 CD11c MACS Micro Bead System Miltenyi Biotec Cat# 130-125-835 Naive CD4+ T Cell Isolation Kit, mouse Miltenyi Biotec Cat# 130-104-453 CFSE Thermo Fisher Scientific Cat# 65-0850-84 Recombinant Mouse TGF-beta 1 R&D Cat# 7666-MB-005/CF Animal-Free Recombinant Human IL-2 Peprotech Cat# AF-200-02 Recombinant mouse PD-L1 Fc-Fusion protein Kerafast Cat# Pr00112-1.9 Recombinant human IgG1 Fc control protein R&D Cat# 110-HG-100 Adult brain dissociation kit Miltenyi Biotec Cat# 130-107-677 Fixable Viability Dye eFluorTM 780 Thermo Fisher Scientific Cat# 65-0865-14 SYTOXTM AADvancedTM Thermo Fisher Scientific Cat# S10274 FoxP3/Transcription factor staining buffer set Thermo Fisher Scientific Cat# 00-5523-00 eBioscienceTM Intracellular fixation & permeabilization buffer set Thermo Fisher Scientific Cat# 88-8824-00 True-Phos Buffer Set BioLegend Cat# 425401 Ionomycin Sigma Aldrich Cat# I0634 PMA Sigma Aldrich Cat# P1585 Brefeldin A solution Thermo Fisher Scientific Cat# 00-4506-51 High-Capacity cDNA Reverse Transcription Kit Thermo Fisher Scientific Cat# 4368814 TaqManTM Gene Expression Master Mix Thermo Fisher Scientific Cat# 4369016 Luna Master Mix New England BioLabs Cat# M3003E Critical commercial assays LEGENDplexTM Mouse Inflammation Panel BioLegend Cat# 740446 LEGENDplexTM Mouse Th Panel BioLegend Cat# 740741 Experimental models: Organisms/strains Mouse: Jak1tm1a(EUCOMM)Hmgu EUMMCR MGI:4456720 Mouse: ROSA26::FLPe knock in; B6.129S4-Gt(ROSA)26Sortm1(FLP1)Dym/RainJ The Jackson Laboratory Cat# JAX:009086, RRID:IMSR_JAX:009086 Mouse: CD11c cre; B6.Cg-Tg(Itgax-cre)11Reiz/J mice The Jackson Laboratory Cat# JAX:008068, RRID:IMSR_JAX:008068 (Continued on next page) Cell Reports 38, 110420, February 22, 2022 e3
Article Title: Succinate uptake by T cells suppresses their effector function via inhibition of mitochondrial glucose oxidation.
Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies anti-CD69-APC Biolegend Clone FN50, Cat# 310910; RRID:AB_314845 anti-CD25-BV605 Biolegend Clone BC96, Cat# 302632; RRID:AB_11218989 anti-CD107a-FITC BD Pharmingen Clone H4A3, Cat# 560949; RRID:AB_2033934 anti-IFN-g-FITC Biolegend Clone B27, Cat# 506504; RRID:AB_315437 anti-TNF-a-PE Biolegend Clone Mab11, Cat# 502909; RRID:AB_315261 anti-IFN-g capture (ELISA) Bio-Rad Clone AbD00676, Cat# HCA043; RRID:AB_906045 anti-IFN-g biotinylated (ELISA) Bio-Rad Clone 2503 Cat# HCA044P; RRID:AB_10614712 anti-HIF-1a [H1alpha67] (WB) Abcam Cat# ab1; RRID:AB_296474 anti-SUCNR1 (WB) Novus Biologicals Cat# NBP1-00861; RRID:AB_1503315 anti-CD3 (mouse) Biolegend Clone 145-2C11 Cat# 100340; RRID:AB_11149115 anti-CD28 (mouse) Biolegend Clone 37.51 Cat# 102116; RRID:AB_11147170 anti-IFN-g-FITC (mouse) Biolegend Clone XMG1.2, Cat# 505806; RRID:AB_315400 anti-TNF-a-PE (mouse) Biolegend Clone MP6-XT22, Cat# 506305; RRID:AB_315426 Chemicals, peptides, and recombinant proteins RPMI1640 Cell Culture Medium Sigma Aldrich Cat# F9665 Penicillin/Streptomycin Thermo Fisher Cat# 15140122 Recombinant human IL-2 PeproTech Cat# 200-02 Immunocult Human CD3/CD28 T cell activator Stemcell Technologies Cat# 10991 Sodium Succinate Sigma Aldrich Cat# 14160 AZD-3965 Selleckchem Cat# S7339 b-chloro-L-alanine Sigma Aldrich Cat# C9033 Sodium Pyruvate Sigma Aldrich Cat# P8574 Dichloroacetate Sigma Aldrich Cat# 347795 Sodium Metavanadate Sigma Aldrich Cat# 72060 Cell Activation Cocktail Biolegend Cat# 423304 Fixation/permeabilization solutions eBioscience Cat# 00-5523-00 CellTrace Violet Thermo Fisher Cat# C34557 Recombinant IFN-g standard (ELISA) Bio-Rad Cat# PHP050 streptavidin-HRP Sigma Aldrich Cat# E2866 TMB substrate Bio-Rad Cat# BUF062B MitoSpy Orange Biolegend Cat# 424804 Laemmli buffer Sigma Aldrich Cat# 38733 RIPA buffer Thermo Fisher Scientific Cat# 89900 Clarity Western ECL Substrate Bio-Rad Cat# 170-5061 U-13C6 glucose CK Isotopes Cat# CLM-1396-1 13C4 sodium succinate Sigma Aldrich Cat# 491985 Methoxyamine hydrochloride in pyridine Thermo Fisher Scientific Cat# 25104 Lipofectamine RNAiMAX Thermo Fisher Cat# 13778075 Fixable Viability Dye eFluorTM 780 Thermo Fisher Scientific Cat# 65-0865-14 4C (SUCNR1 antagonist) Provided by Prof S Pluchino N/A (Continued on next page) Cell Reports 40, 111193, August 16, 2022 e1
Recombinant:Article Title: JAK1 signaling in dendritic cells promotes peripheral tolerance in autoimmunity through PD-L1-mediated regulatory T cell induction.
Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Chemicals, peptides, and recombinant proteins Myelin oligodendrocyte glycoprotein (MOG)35–55 Charitè Berlin NA Incomplete Freund’s adjuvant Sigma-Aldrich Cat# F5506 Mycobacterium tuberculosis H37Ra BD Cat# 231141 Pertussis toxin List Biological Laboratories Cat# 181 recombinant murine GM-CSF R&D Cat# 415-ML-050 recombinant murine IL-4 Miltenyi Biotec Cat# 130-097-761 recombinant murine IFN-ɣ Miltenyi Biotec Cat# 130-105-774 recombinant murine IL-10 R&D Cat# 417-ML-005 Lipopolysaccharide ultra pure InvivoGen Cat# tlrl-pelps Filgotinib TargetMol Cat# T1929 Stattic MedChem Express Cat# HY-13818 Fludarabine MedChem Express Cat# HY-B0069 Ruxolitinib Cayman Chemical Cat# 11609-5 Tofacitinib MedChem Express Cat# HY-40354 OVA323–399 Bachem Cat# H-5532 CD11c MACS Micro Bead System Miltenyi Biotec Cat# 130-125-835 Naive CD4+ T Cell Isolation Kit, mouse Miltenyi Biotec Cat# 130-104-453 CFSE Thermo Fisher Scientific Cat# 65-0850-84 Recombinant Mouse TGF-beta 1 R&D Cat# 7666-MB-005/CF Animal-Free Recombinant Human IL-2 Peprotech Cat# AF-200-02 Recombinant mouse PD-L1 Fc-Fusion protein Kerafast Cat# Pr00112-1.9 Recombinant human IgG1 Fc control protein R&D Cat# 110-HG-100 Adult brain dissociation kit Miltenyi Biotec Cat# 130-107-677 Fixable Viability Dye eFluorTM 780 Thermo Fisher Scientific Cat# 65-0865-14 SYTOXTM AADvancedTM Thermo Fisher Scientific Cat# S10274 FoxP3/Transcription factor staining buffer set Thermo Fisher Scientific Cat# 00-5523-00 eBioscienceTM Intracellular fixation & permeabilization buffer set Thermo Fisher Scientific Cat# 88-8824-00 True-Phos Buffer Set BioLegend Cat# 425401 Ionomycin Sigma Aldrich Cat# I0634 PMA Sigma Aldrich Cat# P1585 Brefeldin A solution Thermo Fisher Scientific Cat# 00-4506-51 High-Capacity cDNA Reverse Transcription Kit Thermo Fisher Scientific Cat# 4368814 TaqManTM Gene Expression Master Mix Thermo Fisher Scientific Cat# 4369016 Luna Master Mix New England BioLabs Cat# M3003E Critical commercial assays LEGENDplexTM Mouse Inflammation Panel BioLegend Cat# 740446 LEGENDplexTM Mouse Th Panel BioLegend Cat# 740741 Experimental models: Organisms/strains Mouse: Jak1tm1a(EUCOMM)Hmgu EUMMCR MGI:4456720 Mouse: ROSA26::FLPe knock in; B6.129S4-Gt(ROSA)26Sortm1(FLP1)Dym/RainJ The Jackson Laboratory Cat# JAX:009086, RRID:IMSR_JAX:009086 Mouse: CD11c cre; B6.Cg-Tg(Itgax-cre)11Reiz/J mice The Jackson Laboratory Cat# JAX:008068, RRID:IMSR_JAX:008068 (Continued on next page) Cell Reports 38, 110420, February 22, 2022 e3
Article Title: Succinate uptake by T cells suppresses their effector function via inhibition of mitochondrial glucose oxidation.
Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies anti-CD69-APC Biolegend Clone FN50, Cat# 310910; RRID:AB_314845 anti-CD25-BV605 Biolegend Clone BC96, Cat# 302632; RRID:AB_11218989 anti-CD107a-FITC BD Pharmingen Clone H4A3, Cat# 560949; RRID:AB_2033934 anti-IFN-g-FITC Biolegend Clone B27, Cat# 506504; RRID:AB_315437 anti-TNF-a-PE Biolegend Clone Mab11, Cat# 502909; RRID:AB_315261 anti-IFN-g capture (ELISA) Bio-Rad Clone AbD00676, Cat# HCA043; RRID:AB_906045 anti-IFN-g biotinylated (ELISA) Bio-Rad Clone 2503 Cat# HCA044P; RRID:AB_10614712 anti-HIF-1a [H1alpha67] (WB) Abcam Cat# ab1; RRID:AB_296474 anti-SUCNR1 (WB) Novus Biologicals Cat# NBP1-00861; RRID:AB_1503315 anti-CD3 (mouse) Biolegend Clone 145-2C11 Cat# 100340; RRID:AB_11149115 anti-CD28 (mouse) Biolegend Clone 37.51 Cat# 102116; RRID:AB_11147170 anti-IFN-g-FITC (mouse) Biolegend Clone XMG1.2, Cat# 505806; RRID:AB_315400 anti-TNF-a-PE (mouse) Biolegend Clone MP6-XT22, Cat# 506305; RRID:AB_315426 Chemicals, peptides, and recombinant proteins RPMI1640 Cell Culture Medium Sigma Aldrich Cat# F9665 Penicillin/Streptomycin Thermo Fisher Cat# 15140122 Recombinant human IL-2 PeproTech Cat# 200-02 Immunocult Human CD3/CD28 T cell activator Stemcell Technologies Cat# 10991 Sodium Succinate Sigma Aldrich Cat# 14160 AZD-3965 Selleckchem Cat# S7339 b-chloro-L-alanine Sigma Aldrich Cat# C9033 Sodium Pyruvate Sigma Aldrich Cat# P8574 Dichloroacetate Sigma Aldrich Cat# 347795 Sodium Metavanadate Sigma Aldrich Cat# 72060 Cell Activation Cocktail Biolegend Cat# 423304 Fixation/permeabilization solutions eBioscience Cat# 00-5523-00 CellTrace Violet Thermo Fisher Cat# C34557 Recombinant IFN-g standard (ELISA) Bio-Rad Cat# PHP050 streptavidin-HRP Sigma Aldrich Cat# E2866 TMB substrate Bio-Rad Cat# BUF062B MitoSpy Orange Biolegend Cat# 424804 Laemmli buffer Sigma Aldrich Cat# 38733 RIPA buffer Thermo Fisher Scientific Cat# 89900 Clarity Western ECL Substrate Bio-Rad Cat# 170-5061 U-13C6 glucose CK Isotopes Cat# CLM-1396-1 13C4 sodium succinate Sigma Aldrich Cat# 491985 Methoxyamine hydrochloride in pyridine Thermo Fisher Scientific Cat# 25104 Lipofectamine RNAiMAX Thermo Fisher Cat# 13778075 Fixable Viability Dye eFluorTM 780 Thermo Fisher Scientific Cat# 65-0865-14 4C (SUCNR1 antagonist) Provided by Prof S Pluchino N/A (Continued on next page) Cell Reports 40, 111193, August 16, 2022 e1
Cell Culture:Article Title: JAK1 signaling in dendritic cells promotes peripheral tolerance in autoimmunity through PD-L1-mediated regulatory T cell induction.
Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Chemicals, peptides, and recombinant proteins Myelin oligodendrocyte glycoprotein (MOG)35–55 Charitè Berlin NA Incomplete Freund’s adjuvant Sigma-Aldrich Cat# F5506 Mycobacterium tuberculosis H37Ra BD Cat# 231141 Pertussis toxin List Biological Laboratories Cat# 181 recombinant murine GM-CSF R&D Cat# 415-ML-050 recombinant murine IL-4 Miltenyi Biotec Cat# 130-097-761 recombinant murine IFN-ɣ Miltenyi Biotec Cat# 130-105-774 recombinant murine IL-10 R&D Cat# 417-ML-005 Lipopolysaccharide ultra pure InvivoGen Cat# tlrl-pelps Filgotinib TargetMol Cat# T1929 Stattic MedChem Express Cat# HY-13818 Fludarabine MedChem Express Cat# HY-B0069 Ruxolitinib Cayman Chemical Cat# 11609-5 Tofacitinib MedChem Express Cat# HY-40354 OVA323–399 Bachem Cat# H-5532 CD11c MACS Micro Bead System Miltenyi Biotec Cat# 130-125-835 Naive CD4+ T Cell Isolation Kit, mouse Miltenyi Biotec Cat# 130-104-453 CFSE Thermo Fisher Scientific Cat# 65-0850-84 Recombinant Mouse TGF-beta 1 R&D Cat# 7666-MB-005/CF Animal-Free Recombinant Human IL-2 Peprotech Cat# AF-200-02 Recombinant mouse PD-L1 Fc-Fusion protein Kerafast Cat# Pr00112-1.9 Recombinant human IgG1 Fc control protein R&D Cat# 110-HG-100 Adult brain dissociation kit Miltenyi Biotec Cat# 130-107-677 Fixable Viability Dye eFluorTM 780 Thermo Fisher Scientific Cat# 65-0865-14 SYTOXTM AADvancedTM Thermo Fisher Scientific Cat# S10274 FoxP3/Transcription factor staining buffer set Thermo Fisher Scientific Cat# 00-5523-00 eBioscienceTM Intracellular fixation & permeabilization buffer set Thermo Fisher Scientific Cat# 88-8824-00 True-Phos Buffer Set BioLegend Cat# 425401 Ionomycin Sigma Aldrich Cat# I0634 PMA Sigma Aldrich Cat# P1585 Brefeldin A solution Thermo Fisher Scientific Cat# 00-4506-51 High-Capacity cDNA Reverse Transcription Kit Thermo Fisher Scientific Cat# 4368814 TaqManTM Gene Expression Master Mix Thermo Fisher Scientific Cat# 4369016 Luna Master Mix New England BioLabs Cat# M3003E Critical commercial assays LEGENDplexTM Mouse Inflammation Panel BioLegend Cat# 740446 LEGENDplexTM Mouse Th Panel BioLegend Cat# 740741 Experimental models: Organisms/strains Mouse: Jak1tm1a(EUCOMM)Hmgu EUMMCR MGI:4456720 Mouse: ROSA26::FLPe knock in; B6.129S4-Gt(ROSA)26Sortm1(FLP1)Dym/RainJ The Jackson Laboratory Cat# JAX:009086, RRID:IMSR_JAX:009086 Mouse: CD11c cre; B6.Cg-Tg(Itgax-cre)11Reiz/J mice The Jackson Laboratory Cat# JAX:008068, RRID:IMSR_JAX:008068 (Continued on next page) Cell Reports 38, 110420, February 22, 2022 e3
Article Title: Succinate uptake by T cells suppresses their effector function via inhibition of mitochondrial glucose oxidation.
Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies anti-CD69-APC Biolegend Clone FN50, Cat# 310910; RRID:AB_314845 anti-CD25-BV605 Biolegend Clone BC96, Cat# 302632; RRID:AB_11218989 anti-CD107a-FITC BD Pharmingen Clone H4A3, Cat# 560949; RRID:AB_2033934 anti-IFN-g-FITC Biolegend Clone B27, Cat# 506504; RRID:AB_315437 anti-TNF-a-PE Biolegend Clone Mab11, Cat# 502909; RRID:AB_315261 anti-IFN-g capture (ELISA) Bio-Rad Clone AbD00676, Cat# HCA043; RRID:AB_906045 anti-IFN-g biotinylated (ELISA) Bio-Rad Clone 2503 Cat# HCA044P; RRID:AB_10614712 anti-HIF-1a [H1alpha67] (WB) Abcam Cat# ab1; RRID:AB_296474 anti-SUCNR1 (WB) Novus Biologicals Cat# NBP1-00861; RRID:AB_1503315 anti-CD3 (mouse) Biolegend Clone 145-2C11 Cat# 100340; RRID:AB_11149115 anti-CD28 (mouse) Biolegend Clone 37.51 Cat# 102116; RRID:AB_11147170 anti-IFN-g-FITC (mouse) Biolegend Clone XMG1.2, Cat# 505806; RRID:AB_315400 anti-TNF-a-PE (mouse) Biolegend Clone MP6-XT22, Cat# 506305; RRID:AB_315426 Chemicals, peptides, and recombinant proteins RPMI1640 Cell Culture Medium Sigma Aldrich Cat# F9665 Penicillin/Streptomycin Thermo Fisher Cat# 15140122 Recombinant human IL-2 PeproTech Cat# 200-02 Immunocult Human CD3/CD28 T cell activator Stemcell Technologies Cat# 10991 Sodium Succinate Sigma Aldrich Cat# 14160 AZD-3965 Selleckchem Cat# S7339 b-chloro-L-alanine Sigma Aldrich Cat# C9033 Sodium Pyruvate Sigma Aldrich Cat# P8574 Dichloroacetate Sigma Aldrich Cat# 347795 Sodium Metavanadate Sigma Aldrich Cat# 72060 Cell Activation Cocktail Biolegend Cat# 423304 Fixation/permeabilization solutions eBioscience Cat# 00-5523-00 CellTrace Violet Thermo Fisher Cat# C34557 Recombinant IFN-g standard (ELISA) Bio-Rad Cat# PHP050 streptavidin-HRP Sigma Aldrich Cat# E2866 TMB substrate Bio-Rad Cat# BUF062B MitoSpy Orange Biolegend Cat# 424804 Laemmli buffer Sigma Aldrich Cat# 38733 RIPA buffer Thermo Fisher Scientific Cat# 89900 Clarity Western ECL Substrate Bio-Rad Cat# 170-5061 U-13C6 glucose CK Isotopes Cat# CLM-1396-1 13C4 sodium succinate Sigma Aldrich Cat# 491985 Methoxyamine hydrochloride in pyridine Thermo Fisher Scientific Cat# 25104 Lipofectamine RNAiMAX Thermo Fisher Cat# 13778075 Fixable Viability Dye eFluorTM 780 Thermo Fisher Scientific Cat# 65-0865-14 4C (SUCNR1 antagonist) Provided by Prof S Pluchino N/A (Continued on next page) Cell Reports 40, 111193, August 16, 2022 e1
Activation Assay:Article Title: JAK1 signaling in dendritic cells promotes peripheral tolerance in autoimmunity through PD-L1-mediated regulatory T cell induction.
Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Chemicals, peptides, and recombinant proteins Myelin oligodendrocyte glycoprotein (MOG)35–55 Charitè Berlin NA Incomplete Freund’s adjuvant Sigma-Aldrich Cat# F5506 Mycobacterium tuberculosis H37Ra BD Cat# 231141 Pertussis toxin List Biological Laboratories Cat# 181 recombinant murine GM-CSF R&D Cat# 415-ML-050 recombinant murine IL-4 Miltenyi Biotec Cat# 130-097-761 recombinant murine IFN-ɣ Miltenyi Biotec Cat# 130-105-774 recombinant murine IL-10 R&D Cat# 417-ML-005 Lipopolysaccharide ultra pure InvivoGen Cat# tlrl-pelps Filgotinib TargetMol Cat# T1929 Stattic MedChem Express Cat# HY-13818 Fludarabine MedChem Express Cat# HY-B0069 Ruxolitinib Cayman Chemical Cat# 11609-5 Tofacitinib MedChem Express Cat# HY-40354 OVA323–399 Bachem Cat# H-5532 CD11c MACS Micro Bead System Miltenyi Biotec Cat# 130-125-835 Naive CD4+ T Cell Isolation Kit, mouse Miltenyi Biotec Cat# 130-104-453 CFSE Thermo Fisher Scientific Cat# 65-0850-84 Recombinant Mouse TGF-beta 1 R&D Cat# 7666-MB-005/CF Animal-Free Recombinant Human IL-2 Peprotech Cat# AF-200-02 Recombinant mouse PD-L1 Fc-Fusion protein Kerafast Cat# Pr00112-1.9 Recombinant human IgG1 Fc control protein R&D Cat# 110-HG-100 Adult brain dissociation kit Miltenyi Biotec Cat# 130-107-677 Fixable Viability Dye eFluorTM 780 Thermo Fisher Scientific Cat# 65-0865-14 SYTOXTM AADvancedTM Thermo Fisher Scientific Cat# S10274 FoxP3/Transcription factor staining buffer set Thermo Fisher Scientific Cat# 00-5523-00 eBioscienceTM Intracellular fixation & permeabilization buffer set Thermo Fisher Scientific Cat# 88-8824-00 True-Phos Buffer Set BioLegend Cat# 425401 Ionomycin Sigma Aldrich Cat# I0634 PMA Sigma Aldrich Cat# P1585 Brefeldin A solution Thermo Fisher Scientific Cat# 00-4506-51 High-Capacity cDNA Reverse Transcription Kit Thermo Fisher Scientific Cat# 4368814 TaqManTM Gene Expression Master Mix Thermo Fisher Scientific Cat# 4369016 Luna Master Mix New England BioLabs Cat# M3003E Critical commercial assays LEGENDplexTM Mouse Inflammation Panel BioLegend Cat# 740446 LEGENDplexTM Mouse Th Panel BioLegend Cat# 740741 Experimental models: Organisms/strains Mouse: Jak1tm1a(EUCOMM)Hmgu EUMMCR MGI:4456720 Mouse: ROSA26::FLPe knock in; B6.129S4-Gt(ROSA)26Sortm1(FLP1)Dym/RainJ The Jackson Laboratory Cat# JAX:009086, RRID:IMSR_JAX:009086 Mouse: CD11c cre; B6.Cg-Tg(Itgax-cre)11Reiz/J mice The Jackson Laboratory Cat# JAX:008068, RRID:IMSR_JAX:008068 (Continued on next page) Cell Reports 38, 110420, February 22, 2022 e3
Article Title: Succinate uptake by T cells suppresses their effector function via inhibition of mitochondrial glucose oxidation.
Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies anti-CD69-APC Biolegend Clone FN50, Cat# 310910; RRID:AB_314845 anti-CD25-BV605 Biolegend Clone BC96, Cat# 302632; RRID:AB_11218989 anti-CD107a-FITC BD Pharmingen Clone H4A3, Cat# 560949; RRID:AB_2033934 anti-IFN-g-FITC Biolegend Clone B27, Cat# 506504; RRID:AB_315437 anti-TNF-a-PE Biolegend Clone Mab11, Cat# 502909; RRID:AB_315261 anti-IFN-g capture (ELISA) Bio-Rad Clone AbD00676, Cat# HCA043; RRID:AB_906045 anti-IFN-g biotinylated (ELISA) Bio-Rad Clone 2503 Cat# HCA044P; RRID:AB_10614712 anti-HIF-1a [H1alpha67] (WB) Abcam Cat# ab1; RRID:AB_296474 anti-SUCNR1 (WB) Novus Biologicals Cat# NBP1-00861; RRID:AB_1503315 anti-CD3 (mouse) Biolegend Clone 145-2C11 Cat# 100340; RRID:AB_11149115 anti-CD28 (mouse) Biolegend Clone 37.51 Cat# 102116; RRID:AB_11147170 anti-IFN-g-FITC (mouse) Biolegend Clone XMG1.2, Cat# 505806; RRID:AB_315400 anti-TNF-a-PE (mouse) Biolegend Clone MP6-XT22, Cat# 506305; RRID:AB_315426 Chemicals, peptides, and recombinant proteins RPMI1640 Cell Culture Medium Sigma Aldrich Cat# F9665 Penicillin/Streptomycin Thermo Fisher Cat# 15140122 Recombinant human IL-2 PeproTech Cat# 200-02 Immunocult Human CD3/CD28 T cell activator Stemcell Technologies Cat# 10991 Sodium Succinate Sigma Aldrich Cat# 14160 AZD-3965 Selleckchem Cat# S7339 b-chloro-L-alanine Sigma Aldrich Cat# C9033 Sodium Pyruvate Sigma Aldrich Cat# P8574 Dichloroacetate Sigma Aldrich Cat# 347795 Sodium Metavanadate Sigma Aldrich Cat# 72060 Cell Activation Cocktail Biolegend Cat# 423304 Fixation/permeabilization solutions eBioscience Cat# 00-5523-00 CellTrace Violet Thermo Fisher Cat# C34557 Recombinant IFN-g standard (ELISA) Bio-Rad Cat# PHP050 streptavidin-HRP Sigma Aldrich Cat# E2866 TMB substrate Bio-Rad Cat# BUF062B MitoSpy Orange Biolegend Cat# 424804 Laemmli buffer Sigma Aldrich Cat# 38733 RIPA buffer Thermo Fisher Scientific Cat# 89900 Clarity Western ECL Substrate Bio-Rad Cat# 170-5061 U-13C6 glucose CK Isotopes Cat# CLM-1396-1 13C4 sodium succinate Sigma Aldrich Cat# 491985 Methoxyamine hydrochloride in pyridine Thermo Fisher Scientific Cat# 25104 Lipofectamine RNAiMAX Thermo Fisher Cat# 13778075 Fixable Viability Dye eFluorTM 780 Thermo Fisher Scientific Cat# 65-0865-14 4C (SUCNR1 antagonist) Provided by Prof S Pluchino N/A (Continued on next page) Cell Reports 40, 111193, August 16, 2022 e1
Adjuvant:Article Title: JAK1 signaling in dendritic cells promotes peripheral tolerance in autoimmunity through PD-L1-mediated regulatory T cell induction.
Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Chemicals, peptides, and recombinant proteins Myelin oligodendrocyte glycoprotein (MOG)35–55 Charitè Berlin NA Incomplete Freund’s adjuvant Sigma-Aldrich Cat# F5506 Mycobacterium tuberculosis H37Ra BD Cat# 231141 Pertussis toxin List Biological Laboratories Cat# 181 recombinant murine GM-CSF R&D Cat# 415-ML-050 recombinant murine IL-4 Miltenyi Biotec Cat# 130-097-761 recombinant murine IFN-ɣ Miltenyi Biotec Cat# 130-105-774 recombinant murine IL-10 R&D Cat# 417-ML-005 Lipopolysaccharide ultra pure InvivoGen Cat# tlrl-pelps Filgotinib TargetMol Cat# T1929 Stattic MedChem Express Cat# HY-13818 Fludarabine MedChem Express Cat# HY-B0069 Ruxolitinib Cayman Chemical Cat# 11609-5 Tofacitinib MedChem Express Cat# HY-40354 OVA323–399 Bachem Cat# H-5532 CD11c MACS Micro Bead System Miltenyi Biotec Cat# 130-125-835 Naive CD4+ T Cell Isolation Kit, mouse Miltenyi Biotec Cat# 130-104-453 CFSE Thermo Fisher Scientific Cat# 65-0850-84 Recombinant Mouse TGF-beta 1 R&D Cat# 7666-MB-005/CF Animal-Free Recombinant Human IL-2 Peprotech Cat# AF-200-02 Recombinant mouse PD-L1 Fc-Fusion protein Kerafast Cat# Pr00112-1.9 Recombinant human IgG1 Fc control protein R&D Cat# 110-HG-100 Adult brain dissociation kit Miltenyi Biotec Cat# 130-107-677 Fixable Viability Dye eFluorTM 780 Thermo Fisher Scientific Cat# 65-0865-14 SYTOXTM AADvancedTM Thermo Fisher Scientific Cat# S10274 FoxP3/Transcription factor staining buffer set Thermo Fisher Scientific Cat# 00-5523-00 eBioscienceTM Intracellular fixation & permeabilization buffer set Thermo Fisher Scientific Cat# 88-8824-00 True-Phos Buffer Set BioLegend Cat# 425401 Ionomycin Sigma Aldrich Cat# I0634 PMA Sigma Aldrich Cat# P1585 Brefeldin A solution Thermo Fisher Scientific Cat# 00-4506-51 High-Capacity cDNA Reverse Transcription Kit Thermo Fisher Scientific Cat# 4368814 TaqManTM Gene Expression Master Mix Thermo Fisher Scientific Cat# 4369016 Luna Master Mix New England BioLabs Cat# M3003E Critical commercial assays LEGENDplexTM Mouse Inflammation Panel BioLegend Cat# 740446 LEGENDplexTM Mouse Th Panel BioLegend Cat# 740741 Experimental models: Organisms/strains Mouse: Jak1tm1a(EUCOMM)Hmgu EUMMCR MGI:4456720 Mouse: ROSA26::FLPe knock in; B6.129S4-Gt(ROSA)26Sortm1(FLP1)Dym/RainJ The Jackson Laboratory Cat# JAX:009086, RRID:IMSR_JAX:009086 Mouse: CD11c cre; B6.Cg-Tg(Itgax-cre)11Reiz/J mice The Jackson Laboratory Cat# JAX:008068, RRID:IMSR_JAX:008068 (Continued on next page) Cell Reports 38, 110420, February 22, 2022 e3
Article Title: Succinate uptake by T cells suppresses their effector function via inhibition of mitochondrial glucose oxidation.
Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies anti-CD69-APC Biolegend Clone FN50, Cat# 310910; RRID:AB_314845 anti-CD25-BV605 Biolegend Clone BC96, Cat# 302632; RRID:AB_11218989 anti-CD107a-FITC BD Pharmingen Clone H4A3, Cat# 560949; RRID:AB_2033934 anti-IFN-g-FITC Biolegend Clone B27, Cat# 506504; RRID:AB_315437 anti-TNF-a-PE Biolegend Clone Mab11, Cat# 502909; RRID:AB_315261 anti-IFN-g capture (ELISA) Bio-Rad Clone AbD00676, Cat# HCA043; RRID:AB_906045 anti-IFN-g biotinylated (ELISA) Bio-Rad Clone 2503 Cat# HCA044P; RRID:AB_10614712 anti-HIF-1a [H1alpha67] (WB) Abcam Cat# ab1; RRID:AB_296474 anti-SUCNR1 (WB) Novus Biologicals Cat# NBP1-00861; RRID:AB_1503315 anti-CD3 (mouse) Biolegend Clone 145-2C11 Cat# 100340; RRID:AB_11149115 anti-CD28 (mouse) Biolegend Clone 37.51 Cat# 102116; RRID:AB_11147170 anti-IFN-g-FITC (mouse) Biolegend Clone XMG1.2, Cat# 505806; RRID:AB_315400 anti-TNF-a-PE (mouse) Biolegend Clone MP6-XT22, Cat# 506305; RRID:AB_315426 Chemicals, peptides, and recombinant proteins RPMI1640 Cell Culture Medium Sigma Aldrich Cat# F9665 Penicillin/Streptomycin Thermo Fisher Cat# 15140122 Recombinant human IL-2 PeproTech Cat# 200-02 Immunocult Human CD3/CD28 T cell activator Stemcell Technologies Cat# 10991 Sodium Succinate Sigma Aldrich Cat# 14160 AZD-3965 Selleckchem Cat# S7339 b-chloro-L-alanine Sigma Aldrich Cat# C9033 Sodium Pyruvate Sigma Aldrich Cat# P8574 Dichloroacetate Sigma Aldrich Cat# 347795 Sodium Metavanadate Sigma Aldrich Cat# 72060 Cell Activation Cocktail Biolegend Cat# 423304 Fixation/permeabilization solutions eBioscience Cat# 00-5523-00 CellTrace Violet Thermo Fisher Cat# C34557 Recombinant IFN-g standard (ELISA) Bio-Rad Cat# PHP050 streptavidin-HRP Sigma Aldrich Cat# E2866 TMB substrate Bio-Rad Cat# BUF062B MitoSpy Orange Biolegend Cat# 424804 Laemmli buffer Sigma Aldrich Cat# 38733 RIPA buffer Thermo Fisher Scientific Cat# 89900 Clarity Western ECL Substrate Bio-Rad Cat# 170-5061 U-13C6 glucose CK Isotopes Cat# CLM-1396-1 13C4 sodium succinate Sigma Aldrich Cat# 491985 Methoxyamine hydrochloride in pyridine Thermo Fisher Scientific Cat# 25104 Lipofectamine RNAiMAX Thermo Fisher Cat# 13778075 Fixable Viability Dye eFluorTM 780 Thermo Fisher Scientific Cat# 65-0865-14 4C (SUCNR1 antagonist) Provided by Prof S Pluchino N/A (Continued on next page) Cell Reports 40, 111193, August 16, 2022 e1
Magnetic Cell Separation:Article Title: JAK1 signaling in dendritic cells promotes peripheral tolerance in autoimmunity through PD-L1-mediated regulatory T cell induction.
Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Chemicals, peptides, and recombinant proteins Myelin oligodendrocyte glycoprotein (MOG)35–55 Charitè Berlin NA Incomplete Freund’s adjuvant Sigma-Aldrich Cat# F5506 Mycobacterium tuberculosis H37Ra BD Cat# 231141 Pertussis toxin List Biological Laboratories Cat# 181 recombinant murine GM-CSF R&D Cat# 415-ML-050 recombinant murine IL-4 Miltenyi Biotec Cat# 130-097-761 recombinant murine IFN-ɣ Miltenyi Biotec Cat# 130-105-774 recombinant murine IL-10 R&D Cat# 417-ML-005 Lipopolysaccharide ultra pure InvivoGen Cat# tlrl-pelps Filgotinib TargetMol Cat# T1929 Stattic MedChem Express Cat# HY-13818 Fludarabine MedChem Express Cat# HY-B0069 Ruxolitinib Cayman Chemical Cat# 11609-5 Tofacitinib MedChem Express Cat# HY-40354 OVA323–399 Bachem Cat# H-5532 CD11c MACS Micro Bead System Miltenyi Biotec Cat# 130-125-835 Naive CD4+ T Cell Isolation Kit, mouse Miltenyi Biotec Cat# 130-104-453 CFSE Thermo Fisher Scientific Cat# 65-0850-84 Recombinant Mouse TGF-beta 1 R&D Cat# 7666-MB-005/CF Animal-Free Recombinant Human IL-2 Peprotech Cat# AF-200-02 Recombinant mouse PD-L1 Fc-Fusion protein Kerafast Cat# Pr00112-1.9 Recombinant human IgG1 Fc control protein R&D Cat# 110-HG-100 Adult brain dissociation kit Miltenyi Biotec Cat# 130-107-677 Fixable Viability Dye eFluorTM 780 Thermo Fisher Scientific Cat# 65-0865-14 SYTOXTM AADvancedTM Thermo Fisher Scientific Cat# S10274 FoxP3/Transcription factor staining buffer set Thermo Fisher Scientific Cat# 00-5523-00 eBioscienceTM Intracellular fixation & permeabilization buffer set Thermo Fisher Scientific Cat# 88-8824-00 True-Phos Buffer Set BioLegend Cat# 425401 Ionomycin Sigma Aldrich Cat# I0634 PMA Sigma Aldrich Cat# P1585 Brefeldin A solution Thermo Fisher Scientific Cat# 00-4506-51 High-Capacity cDNA Reverse Transcription Kit Thermo Fisher Scientific Cat# 4368814 TaqManTM Gene Expression Master Mix Thermo Fisher Scientific Cat# 4369016 Luna Master Mix New England BioLabs Cat# M3003E Critical commercial assays LEGENDplexTM Mouse Inflammation Panel BioLegend Cat# 740446 LEGENDplexTM Mouse Th Panel BioLegend Cat# 740741 Experimental models: Organisms/strains Mouse: Jak1tm1a(EUCOMM)Hmgu EUMMCR MGI:4456720 Mouse: ROSA26::FLPe knock in; B6.129S4-Gt(ROSA)26Sortm1(FLP1)Dym/RainJ The Jackson Laboratory Cat# JAX:009086, RRID:IMSR_JAX:009086 Mouse: CD11c cre; B6.Cg-Tg(Itgax-cre)11Reiz/J mice The Jackson Laboratory Cat# JAX:008068, RRID:IMSR_JAX:008068 (Continued on next page) Cell Reports 38, 110420, February 22, 2022 e3
Article Title: Succinate uptake by T cells suppresses their effector function via inhibition of mitochondrial glucose oxidation.
Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies anti-CD69-APC Biolegend Clone FN50, Cat# 310910; RRID:AB_314845 anti-CD25-BV605 Biolegend Clone BC96, Cat# 302632; RRID:AB_11218989 anti-CD107a-FITC BD Pharmingen Clone H4A3, Cat# 560949; RRID:AB_2033934 anti-IFN-g-FITC Biolegend Clone B27, Cat# 506504; RRID:AB_315437 anti-TNF-a-PE Biolegend Clone Mab11, Cat# 502909; RRID:AB_315261 anti-IFN-g capture (ELISA) Bio-Rad Clone AbD00676, Cat# HCA043; RRID:AB_906045 anti-IFN-g biotinylated (ELISA) Bio-Rad Clone 2503 Cat# HCA044P; RRID:AB_10614712 anti-HIF-1a [H1alpha67] (WB) Abcam Cat# ab1; RRID:AB_296474 anti-SUCNR1 (WB) Novus Biologicals Cat# NBP1-00861; RRID:AB_1503315 anti-CD3 (mouse) Biolegend Clone 145-2C11 Cat# 100340; RRID:AB_11149115 anti-CD28 (mouse) Biolegend Clone 37.51 Cat# 102116; RRID:AB_11147170 anti-IFN-g-FITC (mouse) Biolegend Clone XMG1.2, Cat# 505806; RRID:AB_315400 anti-TNF-a-PE (mouse) Biolegend Clone MP6-XT22, Cat# 506305; RRID:AB_315426 Chemicals, peptides, and recombinant proteins RPMI1640 Cell Culture Medium Sigma Aldrich Cat# F9665 Penicillin/Streptomycin Thermo Fisher Cat# 15140122 Recombinant human IL-2 PeproTech Cat# 200-02 Immunocult Human CD3/CD28 T cell activator Stemcell Technologies Cat# 10991 Sodium Succinate Sigma Aldrich Cat# 14160 AZD-3965 Selleckchem Cat# S7339 b-chloro-L-alanine Sigma Aldrich Cat# C9033 Sodium Pyruvate Sigma Aldrich Cat# P8574 Dichloroacetate Sigma Aldrich Cat# 347795 Sodium Metavanadate Sigma Aldrich Cat# 72060 Cell Activation Cocktail Biolegend Cat# 423304 Fixation/permeabilization solutions eBioscience Cat# 00-5523-00 CellTrace Violet Thermo Fisher Cat# C34557 Recombinant IFN-g standard (ELISA) Bio-Rad Cat# PHP050 streptavidin-HRP Sigma Aldrich Cat# E2866 TMB substrate Bio-Rad Cat# BUF062B MitoSpy Orange Biolegend Cat# 424804 Laemmli buffer Sigma Aldrich Cat# 38733 RIPA buffer Thermo Fisher Scientific Cat# 89900 Clarity Western ECL Substrate Bio-Rad Cat# 170-5061 U-13C6 glucose CK Isotopes Cat# CLM-1396-1 13C4 sodium succinate Sigma Aldrich Cat# 491985 Methoxyamine hydrochloride in pyridine Thermo Fisher Scientific Cat# 25104 Lipofectamine RNAiMAX Thermo Fisher Cat# 13778075 Fixable Viability Dye eFluorTM 780 Thermo Fisher Scientific Cat# 65-0865-14 4C (SUCNR1 antagonist) Provided by Prof S Pluchino N/A (Continued on next page) Cell Reports 40, 111193, August 16, 2022 e1
Cell Isolation:Article Title: JAK1 signaling in dendritic cells promotes peripheral tolerance in autoimmunity through PD-L1-mediated regulatory T cell induction.
Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Chemicals, peptides, and recombinant proteins Myelin oligodendrocyte glycoprotein (MOG)35–55 Charitè Berlin NA Incomplete Freund’s adjuvant Sigma-Aldrich Cat# F5506 Mycobacterium tuberculosis H37Ra BD Cat# 231141 Pertussis toxin List Biological Laboratories Cat# 181 recombinant murine GM-CSF R&D Cat# 415-ML-050 recombinant murine IL-4 Miltenyi Biotec Cat# 130-097-761 recombinant murine IFN-ɣ Miltenyi Biotec Cat# 130-105-774 recombinant murine IL-10 R&D Cat# 417-ML-005 Lipopolysaccharide ultra pure InvivoGen Cat# tlrl-pelps Filgotinib TargetMol Cat# T1929 Stattic MedChem Express Cat# HY-13818 Fludarabine MedChem Express Cat# HY-B0069 Ruxolitinib Cayman Chemical Cat# 11609-5 Tofacitinib MedChem Express Cat# HY-40354 OVA323–399 Bachem Cat# H-5532 CD11c MACS Micro Bead System Miltenyi Biotec Cat# 130-125-835 Naive CD4+ T Cell Isolation Kit, mouse Miltenyi Biotec Cat# 130-104-453 CFSE Thermo Fisher Scientific Cat# 65-0850-84 Recombinant Mouse TGF-beta 1 R&D Cat# 7666-MB-005/CF Animal-Free Recombinant Human IL-2 Peprotech Cat# AF-200-02 Recombinant mouse PD-L1 Fc-Fusion protein Kerafast Cat# Pr00112-1.9 Recombinant human IgG1 Fc control protein R&D Cat# 110-HG-100 Adult brain dissociation kit Miltenyi Biotec Cat# 130-107-677 Fixable Viability Dye eFluorTM 780 Thermo Fisher Scientific Cat# 65-0865-14 SYTOXTM AADvancedTM Thermo Fisher Scientific Cat# S10274 FoxP3/Transcription factor staining buffer set Thermo Fisher Scientific Cat# 00-5523-00 eBioscienceTM Intracellular fixation & permeabilization buffer set Thermo Fisher Scientific Cat# 88-8824-00 True-Phos Buffer Set BioLegend Cat# 425401 Ionomycin Sigma Aldrich Cat# I0634 PMA Sigma Aldrich Cat# P1585 Brefeldin A solution Thermo Fisher Scientific Cat# 00-4506-51 High-Capacity cDNA Reverse Transcription Kit Thermo Fisher Scientific Cat# 4368814 TaqManTM Gene Expression Master Mix Thermo Fisher Scientific Cat# 4369016 Luna Master Mix New England BioLabs Cat# M3003E Critical commercial assays LEGENDplexTM Mouse Inflammation Panel BioLegend Cat# 740446 LEGENDplexTM Mouse Th Panel BioLegend Cat# 740741 Experimental models: Organisms/strains Mouse: Jak1tm1a(EUCOMM)Hmgu EUMMCR MGI:4456720 Mouse: ROSA26::FLPe knock in; B6.129S4-Gt(ROSA)26Sortm1(FLP1)Dym/RainJ The Jackson Laboratory Cat# JAX:009086, RRID:IMSR_JAX:009086 Mouse: CD11c cre; B6.Cg-Tg(Itgax-cre)11Reiz/J mice The Jackson Laboratory Cat# JAX:008068, RRID:IMSR_JAX:008068 (Continued on next page) Cell Reports 38, 110420, February 22, 2022 e3
Article Title: Succinate uptake by T cells suppresses their effector function via inhibition of mitochondrial glucose oxidation.
Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies anti-CD69-APC Biolegend Clone FN50, Cat# 310910; RRID:AB_314845 anti-CD25-BV605 Biolegend Clone BC96, Cat# 302632; RRID:AB_11218989 anti-CD107a-FITC BD Pharmingen Clone H4A3, Cat# 560949; RRID:AB_2033934 anti-IFN-g-FITC Biolegend Clone B27, Cat# 506504; RRID:AB_315437 anti-TNF-a-PE Biolegend Clone Mab11, Cat# 502909; RRID:AB_315261 anti-IFN-g capture (ELISA) Bio-Rad Clone AbD00676, Cat# HCA043; RRID:AB_906045 anti-IFN-g biotinylated (ELISA) Bio-Rad Clone 2503 Cat# HCA044P; RRID:AB_10614712 anti-HIF-1a [H1alpha67] (WB) Abcam Cat# ab1; RRID:AB_296474 anti-SUCNR1 (WB) Novus Biologicals Cat# NBP1-00861; RRID:AB_1503315 anti-CD3 (mouse) Biolegend Clone 145-2C11 Cat# 100340; RRID:AB_11149115 anti-CD28 (mouse) Biolegend Clone 37.51 Cat# 102116; RRID:AB_11147170 anti-IFN-g-FITC (mouse) Biolegend Clone XMG1.2, Cat# 505806; RRID:AB_315400 anti-TNF-a-PE (mouse) Biolegend Clone MP6-XT22, Cat# 506305; RRID:AB_315426 Chemicals, peptides, and recombinant proteins RPMI1640 Cell Culture Medium Sigma Aldrich Cat# F9665 Penicillin/Streptomycin Thermo Fisher Cat# 15140122 Recombinant human IL-2 PeproTech Cat# 200-02 Immunocult Human CD3/CD28 T cell activator Stemcell Technologies Cat# 10991 Sodium Succinate Sigma Aldrich Cat# 14160 AZD-3965 Selleckchem Cat# S7339 b-chloro-L-alanine Sigma Aldrich Cat# C9033 Sodium Pyruvate Sigma Aldrich Cat# P8574 Dichloroacetate Sigma Aldrich Cat# 347795 Sodium Metavanadate Sigma Aldrich Cat# 72060 Cell Activation Cocktail Biolegend Cat# 423304 Fixation/permeabilization solutions eBioscience Cat# 00-5523-00 CellTrace Violet Thermo Fisher Cat# C34557 Recombinant IFN-g standard (ELISA) Bio-Rad Cat# PHP050 streptavidin-HRP Sigma Aldrich Cat# E2866 TMB substrate Bio-Rad Cat# BUF062B MitoSpy Orange Biolegend Cat# 424804 Laemmli buffer Sigma Aldrich Cat# 38733 RIPA buffer Thermo Fisher Scientific Cat# 89900 Clarity Western ECL Substrate Bio-Rad Cat# 170-5061 U-13C6 glucose CK Isotopes Cat# CLM-1396-1 13C4 sodium succinate Sigma Aldrich Cat# 491985 Methoxyamine hydrochloride in pyridine Thermo Fisher Scientific Cat# 25104 Lipofectamine RNAiMAX Thermo Fisher Cat# 13778075 Fixable Viability Dye eFluorTM 780 Thermo Fisher Scientific Cat# 65-0865-14 4C (SUCNR1 antagonist) Provided by Prof S Pluchino N/A (Continued on next page) Cell Reports 40, 111193, August 16, 2022 e1
Control:Article Title: JAK1 signaling in dendritic cells promotes peripheral tolerance in autoimmunity through PD-L1-mediated regulatory T cell induction.
Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Chemicals, peptides, and recombinant proteins Myelin oligodendrocyte glycoprotein (MOG)35–55 Charitè Berlin NA Incomplete Freund’s adjuvant Sigma-Aldrich Cat# F5506 Mycobacterium tuberculosis H37Ra BD Cat# 231141 Pertussis toxin List Biological Laboratories Cat# 181 recombinant murine GM-CSF R&D Cat# 415-ML-050 recombinant murine IL-4 Miltenyi Biotec Cat# 130-097-761 recombinant murine IFN-ɣ Miltenyi Biotec Cat# 130-105-774 recombinant murine IL-10 R&D Cat# 417-ML-005 Lipopolysaccharide ultra pure InvivoGen Cat# tlrl-pelps Filgotinib TargetMol Cat# T1929 Stattic MedChem Express Cat# HY-13818 Fludarabine MedChem Express Cat# HY-B0069 Ruxolitinib Cayman Chemical Cat# 11609-5 Tofacitinib MedChem Express Cat# HY-40354 OVA323–399 Bachem Cat# H-5532 CD11c MACS Micro Bead System Miltenyi Biotec Cat# 130-125-835 Naive CD4+ T Cell Isolation Kit, mouse Miltenyi Biotec Cat# 130-104-453 CFSE Thermo Fisher Scientific Cat# 65-0850-84 Recombinant Mouse TGF-beta 1 R&D Cat# 7666-MB-005/CF Animal-Free Recombinant Human IL-2 Peprotech Cat# AF-200-02 Recombinant mouse PD-L1 Fc-Fusion protein Kerafast Cat# Pr00112-1.9 Recombinant human IgG1 Fc control protein R&D Cat# 110-HG-100 Adult brain dissociation kit Miltenyi Biotec Cat# 130-107-677 Fixable Viability Dye eFluorTM 780 Thermo Fisher Scientific Cat# 65-0865-14 SYTOXTM AADvancedTM Thermo Fisher Scientific Cat# S10274 FoxP3/Transcription factor staining buffer set Thermo Fisher Scientific Cat# 00-5523-00 eBioscienceTM Intracellular fixation & permeabilization buffer set Thermo Fisher Scientific Cat# 88-8824-00 True-Phos Buffer Set BioLegend Cat# 425401 Ionomycin Sigma Aldrich Cat# I0634 PMA Sigma Aldrich Cat# P1585 Brefeldin A solution Thermo Fisher Scientific Cat# 00-4506-51 High-Capacity cDNA Reverse Transcription Kit Thermo Fisher Scientific Cat# 4368814 TaqManTM Gene Expression Master Mix Thermo Fisher Scientific Cat# 4369016 Luna Master Mix New England BioLabs Cat# M3003E Critical commercial assays LEGENDplexTM Mouse Inflammation Panel BioLegend Cat# 740446 LEGENDplexTM Mouse Th Panel BioLegend Cat# 740741 Experimental models: Organisms/strains Mouse: Jak1tm1a(EUCOMM)Hmgu EUMMCR MGI:4456720 Mouse: ROSA26::FLPe knock in; B6.129S4-Gt(ROSA)26Sortm1(FLP1)Dym/RainJ The Jackson Laboratory Cat# JAX:009086, RRID:IMSR_JAX:009086 Mouse: CD11c cre; B6.Cg-Tg(Itgax-cre)11Reiz/J mice The Jackson Laboratory Cat# JAX:008068, RRID:IMSR_JAX:008068 (Continued on next page) Cell Reports 38, 110420, February 22, 2022 e3
Article Title: Succinate uptake by T cells suppresses their effector function via inhibition of mitochondrial glucose oxidation.
Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies anti-CD69-APC Biolegend Clone FN50, Cat# 310910; RRID:AB_314845 anti-CD25-BV605 Biolegend Clone BC96, Cat# 302632; RRID:AB_11218989 anti-CD107a-FITC BD Pharmingen Clone H4A3, Cat# 560949; RRID:AB_2033934 anti-IFN-g-FITC Biolegend Clone B27, Cat# 506504; RRID:AB_315437 anti-TNF-a-PE Biolegend Clone Mab11, Cat# 502909; RRID:AB_315261 anti-IFN-g capture (ELISA) Bio-Rad Clone AbD00676, Cat# HCA043; RRID:AB_906045 anti-IFN-g biotinylated (ELISA) Bio-Rad Clone 2503 Cat# HCA044P; RRID:AB_10614712 anti-HIF-1a [H1alpha67] (WB) Abcam Cat# ab1; RRID:AB_296474 anti-SUCNR1 (WB) Novus Biologicals Cat# NBP1-00861; RRID:AB_1503315 anti-CD3 (mouse) Biolegend Clone 145-2C11 Cat# 100340; RRID:AB_11149115 anti-CD28 (mouse) Biolegend Clone 37.51 Cat# 102116; RRID:AB_11147170 anti-IFN-g-FITC (mouse) Biolegend Clone XMG1.2, Cat# 505806; RRID:AB_315400 anti-TNF-a-PE (mouse) Biolegend Clone MP6-XT22, Cat# 506305; RRID:AB_315426 Chemicals, peptides, and recombinant proteins RPMI1640 Cell Culture Medium Sigma Aldrich Cat# F9665 Penicillin/Streptomycin Thermo Fisher Cat# 15140122 Recombinant human IL-2 PeproTech Cat# 200-02 Immunocult Human CD3/CD28 T cell activator Stemcell Technologies Cat# 10991 Sodium Succinate Sigma Aldrich Cat# 14160 AZD-3965 Selleckchem Cat# S7339 b-chloro-L-alanine Sigma Aldrich Cat# C9033 Sodium Pyruvate Sigma Aldrich Cat# P8574 Dichloroacetate Sigma Aldrich Cat# 347795 Sodium Metavanadate Sigma Aldrich Cat# 72060 Cell Activation Cocktail Biolegend Cat# 423304 Fixation/permeabilization solutions eBioscience Cat# 00-5523-00 CellTrace Violet Thermo Fisher Cat# C34557 Recombinant IFN-g standard (ELISA) Bio-Rad Cat# PHP050 streptavidin-HRP Sigma Aldrich Cat# E2866 TMB substrate Bio-Rad Cat# BUF062B MitoSpy Orange Biolegend Cat# 424804 Laemmli buffer Sigma Aldrich Cat# 38733 RIPA buffer Thermo Fisher Scientific Cat# 89900 Clarity Western ECL Substrate Bio-Rad Cat# 170-5061 U-13C6 glucose CK Isotopes Cat# CLM-1396-1 13C4 sodium succinate Sigma Aldrich Cat# 491985 Methoxyamine hydrochloride in pyridine Thermo Fisher Scientific Cat# 25104 Lipofectamine RNAiMAX Thermo Fisher Cat# 13778075 Fixable Viability Dye eFluorTM 780 Thermo Fisher Scientific Cat# 65-0865-14 4C (SUCNR1 antagonist) Provided by Prof S Pluchino N/A (Continued on next page) Cell Reports 40, 111193, August 16, 2022 e1
Staining:Article Title: JAK1 signaling in dendritic cells promotes peripheral tolerance in autoimmunity through PD-L1-mediated regulatory T cell induction.
Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Chemicals, peptides, and recombinant proteins Myelin oligodendrocyte glycoprotein (MOG)35–55 Charitè Berlin NA Incomplete Freund’s adjuvant Sigma-Aldrich Cat# F5506 Mycobacterium tuberculosis H37Ra BD Cat# 231141 Pertussis toxin List Biological Laboratories Cat# 181 recombinant murine GM-CSF R&D Cat# 415-ML-050 recombinant murine IL-4 Miltenyi Biotec Cat# 130-097-761 recombinant murine IFN-ɣ Miltenyi Biotec Cat# 130-105-774 recombinant murine IL-10 R&D Cat# 417-ML-005 Lipopolysaccharide ultra pure InvivoGen Cat# tlrl-pelps Filgotinib TargetMol Cat# T1929 Stattic MedChem Express Cat# HY-13818 Fludarabine MedChem Express Cat# HY-B0069 Ruxolitinib Cayman Chemical Cat# 11609-5 Tofacitinib MedChem Express Cat# HY-40354 OVA323–399 Bachem Cat# H-5532 CD11c MACS Micro Bead System Miltenyi Biotec Cat# 130-125-835 Naive CD4+ T Cell Isolation Kit, mouse Miltenyi Biotec Cat# 130-104-453 CFSE Thermo Fisher Scientific Cat# 65-0850-84 Recombinant Mouse TGF-beta 1 R&D Cat# 7666-MB-005/CF Animal-Free Recombinant Human IL-2 Peprotech Cat# AF-200-02 Recombinant mouse PD-L1 Fc-Fusion protein Kerafast Cat# Pr00112-1.9 Recombinant human IgG1 Fc control protein R&D Cat# 110-HG-100 Adult brain dissociation kit Miltenyi Biotec Cat# 130-107-677 Fixable Viability Dye eFluorTM 780 Thermo Fisher Scientific Cat# 65-0865-14 SYTOXTM AADvancedTM Thermo Fisher Scientific Cat# S10274 FoxP3/Transcription factor staining buffer set Thermo Fisher Scientific Cat# 00-5523-00 eBioscienceTM Intracellular fixation & permeabilization buffer set Thermo Fisher Scientific Cat# 88-8824-00 True-Phos Buffer Set BioLegend Cat# 425401 Ionomycin Sigma Aldrich Cat# I0634 PMA Sigma Aldrich Cat# P1585 Brefeldin A solution Thermo Fisher Scientific Cat# 00-4506-51 High-Capacity cDNA Reverse Transcription Kit Thermo Fisher Scientific Cat# 4368814 TaqManTM Gene Expression Master Mix Thermo Fisher Scientific Cat# 4369016 Luna Master Mix New England BioLabs Cat# M3003E Critical commercial assays LEGENDplexTM Mouse Inflammation Panel BioLegend Cat# 740446 LEGENDplexTM Mouse Th Panel BioLegend Cat# 740741 Experimental models: Organisms/strains Mouse: Jak1tm1a(EUCOMM)Hmgu EUMMCR MGI:4456720 Mouse: ROSA26::FLPe knock in; B6.129S4-Gt(ROSA)26Sortm1(FLP1)Dym/RainJ The Jackson Laboratory Cat# JAX:009086, RRID:IMSR_JAX:009086 Mouse: CD11c cre; B6.Cg-Tg(Itgax-cre)11Reiz/J mice The Jackson Laboratory Cat# JAX:008068, RRID:IMSR_JAX:008068 (Continued on next page) Cell Reports 38, 110420, February 22, 2022 e3
Article Title: Succinate uptake by T cells suppresses their effector function via inhibition of mitochondrial glucose oxidation.
Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies anti-CD69-APC Biolegend Clone FN50, Cat# 310910; RRID:AB_314845 anti-CD25-BV605 Biolegend Clone BC96, Cat# 302632; RRID:AB_11218989 anti-CD107a-FITC BD Pharmingen Clone H4A3, Cat# 560949; RRID:AB_2033934 anti-IFN-g-FITC Biolegend Clone B27, Cat# 506504; RRID:AB_315437 anti-TNF-a-PE Biolegend Clone Mab11, Cat# 502909; RRID:AB_315261 anti-IFN-g capture (ELISA) Bio-Rad Clone AbD00676, Cat# HCA043; RRID:AB_906045 anti-IFN-g biotinylated (ELISA) Bio-Rad Clone 2503 Cat# HCA044P; RRID:AB_10614712 anti-HIF-1a [H1alpha67] (WB) Abcam Cat# ab1; RRID:AB_296474 anti-SUCNR1 (WB) Novus Biologicals Cat# NBP1-00861; RRID:AB_1503315 anti-CD3 (mouse) Biolegend Clone 145-2C11 Cat# 100340; RRID:AB_11149115 anti-CD28 (mouse) Biolegend Clone 37.51 Cat# 102116; RRID:AB_11147170 anti-IFN-g-FITC (mouse) Biolegend Clone XMG1.2, Cat# 505806; RRID:AB_315400 anti-TNF-a-PE (mouse) Biolegend Clone MP6-XT22, Cat# 506305; RRID:AB_315426 Chemicals, peptides, and recombinant proteins RPMI1640 Cell Culture Medium Sigma Aldrich Cat# F9665 Penicillin/Streptomycin Thermo Fisher Cat# 15140122 Recombinant human IL-2 PeproTech Cat# 200-02 Immunocult Human CD3/CD28 T cell activator Stemcell Technologies Cat# 10991 Sodium Succinate Sigma Aldrich Cat# 14160 AZD-3965 Selleckchem Cat# S7339 b-chloro-L-alanine Sigma Aldrich Cat# C9033 Sodium Pyruvate Sigma Aldrich Cat# P8574 Dichloroacetate Sigma Aldrich Cat# 347795 Sodium Metavanadate Sigma Aldrich Cat# 72060 Cell Activation Cocktail Biolegend Cat# 423304 Fixation/permeabilization solutions eBioscience Cat# 00-5523-00 CellTrace Violet Thermo Fisher Cat# C34557 Recombinant IFN-g standard (ELISA) Bio-Rad Cat# PHP050 streptavidin-HRP Sigma Aldrich Cat# E2866 TMB substrate Bio-Rad Cat# BUF062B MitoSpy Orange Biolegend Cat# 424804 Laemmli buffer Sigma Aldrich Cat# 38733 RIPA buffer Thermo Fisher Scientific Cat# 89900 Clarity Western ECL Substrate Bio-Rad Cat# 170-5061 U-13C6 glucose CK Isotopes Cat# CLM-1396-1 13C4 sodium succinate Sigma Aldrich Cat# 491985 Methoxyamine hydrochloride in pyridine Thermo Fisher Scientific Cat# 25104 Lipofectamine RNAiMAX Thermo Fisher Cat# 13778075 Fixable Viability Dye eFluorTM 780 Thermo Fisher Scientific Cat# 65-0865-14 4C (SUCNR1 antagonist) Provided by Prof S Pluchino N/A (Continued on next page) Cell Reports 40, 111193, August 16, 2022 e1
Reverse Transcription:Article Title: JAK1 signaling in dendritic cells promotes peripheral tolerance in autoimmunity through PD-L1-mediated regulatory T cell induction.
Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Chemicals, peptides, and recombinant proteins Myelin oligodendrocyte glycoprotein (MOG)35–55 Charitè Berlin NA Incomplete Freund’s adjuvant Sigma-Aldrich Cat# F5506 Mycobacterium tuberculosis H37Ra BD Cat# 231141 Pertussis toxin List Biological Laboratories Cat# 181 recombinant murine GM-CSF R&D Cat# 415-ML-050 recombinant murine IL-4 Miltenyi Biotec Cat# 130-097-761 recombinant murine IFN-ɣ Miltenyi Biotec Cat# 130-105-774 recombinant murine IL-10 R&D Cat# 417-ML-005 Lipopolysaccharide ultra pure InvivoGen Cat# tlrl-pelps Filgotinib TargetMol Cat# T1929 Stattic MedChem Express Cat# HY-13818 Fludarabine MedChem Express Cat# HY-B0069 Ruxolitinib Cayman Chemical Cat# 11609-5 Tofacitinib MedChem Express Cat# HY-40354 OVA323–399 Bachem Cat# H-5532 CD11c MACS Micro Bead System Miltenyi Biotec Cat# 130-125-835 Naive CD4+ T Cell Isolation Kit, mouse Miltenyi Biotec Cat# 130-104-453 CFSE Thermo Fisher Scientific Cat# 65-0850-84 Recombinant Mouse TGF-beta 1 R&D Cat# 7666-MB-005/CF Animal-Free Recombinant Human IL-2 Peprotech Cat# AF-200-02 Recombinant mouse PD-L1 Fc-Fusion protein Kerafast Cat# Pr00112-1.9 Recombinant human IgG1 Fc control protein R&D Cat# 110-HG-100 Adult brain dissociation kit Miltenyi Biotec Cat# 130-107-677 Fixable Viability Dye eFluorTM 780 Thermo Fisher Scientific Cat# 65-0865-14 SYTOXTM AADvancedTM Thermo Fisher Scientific Cat# S10274 FoxP3/Transcription factor staining buffer set Thermo Fisher Scientific Cat# 00-5523-00 eBioscienceTM Intracellular fixation & permeabilization buffer set Thermo Fisher Scientific Cat# 88-8824-00 True-Phos Buffer Set BioLegend Cat# 425401 Ionomycin Sigma Aldrich Cat# I0634 PMA Sigma Aldrich Cat# P1585 Brefeldin A solution Thermo Fisher Scientific Cat# 00-4506-51 High-Capacity cDNA Reverse Transcription Kit Thermo Fisher Scientific Cat# 4368814 TaqManTM Gene Expression Master Mix Thermo Fisher Scientific Cat# 4369016 Luna Master Mix New England BioLabs Cat# M3003E Critical commercial assays LEGENDplexTM Mouse Inflammation Panel BioLegend Cat# 740446 LEGENDplexTM Mouse Th Panel BioLegend Cat# 740741 Experimental models: Organisms/strains Mouse: Jak1tm1a(EUCOMM)Hmgu EUMMCR MGI:4456720 Mouse: ROSA26::FLPe knock in; B6.129S4-Gt(ROSA)26Sortm1(FLP1)Dym/RainJ The Jackson Laboratory Cat# JAX:009086, RRID:IMSR_JAX:009086 Mouse: CD11c cre; B6.Cg-Tg(Itgax-cre)11Reiz/J mice The Jackson Laboratory Cat# JAX:008068, RRID:IMSR_JAX:008068 (Continued on next page) Cell Reports 38, 110420, February 22, 2022 e3
Article Title: Succinate uptake by T cells suppresses their effector function via inhibition of mitochondrial glucose oxidation.
Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies anti-CD69-APC Biolegend Clone FN50, Cat# 310910; RRID:AB_314845 anti-CD25-BV605 Biolegend Clone BC96, Cat# 302632; RRID:AB_11218989 anti-CD107a-FITC BD Pharmingen Clone H4A3, Cat# 560949; RRID:AB_2033934 anti-IFN-g-FITC Biolegend Clone B27, Cat# 506504; RRID:AB_315437 anti-TNF-a-PE Biolegend Clone Mab11, Cat# 502909; RRID:AB_315261 anti-IFN-g capture (ELISA) Bio-Rad Clone AbD00676, Cat# HCA043; RRID:AB_906045 anti-IFN-g biotinylated (ELISA) Bio-Rad Clone 2503 Cat# HCA044P; RRID:AB_10614712 anti-HIF-1a [H1alpha67] (WB) Abcam Cat# ab1; RRID:AB_296474 anti-SUCNR1 (WB) Novus Biologicals Cat# NBP1-00861; RRID:AB_1503315 anti-CD3 (mouse) Biolegend Clone 145-2C11 Cat# 100340; RRID:AB_11149115 anti-CD28 (mouse) Biolegend Clone 37.51 Cat# 102116; RRID:AB_11147170 anti-IFN-g-FITC (mouse) Biolegend Clone XMG1.2, Cat# 505806; RRID:AB_315400 anti-TNF-a-PE (mouse) Biolegend Clone MP6-XT22, Cat# 506305; RRID:AB_315426 Chemicals, peptides, and recombinant proteins RPMI1640 Cell Culture Medium Sigma Aldrich Cat# F9665 Penicillin/Streptomycin Thermo Fisher Cat# 15140122 Recombinant human IL-2 PeproTech Cat# 200-02 Immunocult Human CD3/CD28 T cell activator Stemcell Technologies Cat# 10991 Sodium Succinate Sigma Aldrich Cat# 14160 AZD-3965 Selleckchem Cat# S7339 b-chloro-L-alanine Sigma Aldrich Cat# C9033 Sodium Pyruvate Sigma Aldrich Cat# P8574 Dichloroacetate Sigma Aldrich Cat# 347795 Sodium Metavanadate Sigma Aldrich Cat# 72060 Cell Activation Cocktail Biolegend Cat# 423304 Fixation/permeabilization solutions eBioscience Cat# 00-5523-00 CellTrace Violet Thermo Fisher Cat# C34557 Recombinant IFN-g standard (ELISA) Bio-Rad Cat# PHP050 streptavidin-HRP Sigma Aldrich Cat# E2866 TMB substrate Bio-Rad Cat# BUF062B MitoSpy Orange Biolegend Cat# 424804 Laemmli buffer Sigma Aldrich Cat# 38733 RIPA buffer Thermo Fisher Scientific Cat# 89900 Clarity Western ECL Substrate Bio-Rad Cat# 170-5061 U-13C6 glucose CK Isotopes Cat# CLM-1396-1 13C4 sodium succinate Sigma Aldrich Cat# 491985 Methoxyamine hydrochloride in pyridine Thermo Fisher Scientific Cat# 25104 Lipofectamine RNAiMAX Thermo Fisher Cat# 13778075 Fixable Viability Dye eFluorTM 780 Thermo Fisher Scientific Cat# 65-0865-14 4C (SUCNR1 antagonist) Provided by Prof S Pluchino N/A (Continued on next page) Cell Reports 40, 111193, August 16, 2022 e1
Expressing:Article Title: JAK1 signaling in dendritic cells promotes peripheral tolerance in autoimmunity through PD-L1-mediated regulatory T cell induction.
Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Chemicals, peptides, and recombinant proteins Myelin oligodendrocyte glycoprotein (MOG)35–55 Charitè Berlin NA Incomplete Freund’s adjuvant Sigma-Aldrich Cat# F5506 Mycobacterium tuberculosis H37Ra BD Cat# 231141 Pertussis toxin List Biological Laboratories Cat# 181 recombinant murine GM-CSF R&D Cat# 415-ML-050 recombinant murine IL-4 Miltenyi Biotec Cat# 130-097-761 recombinant murine IFN-ɣ Miltenyi Biotec Cat# 130-105-774 recombinant murine IL-10 R&D Cat# 417-ML-005 Lipopolysaccharide ultra pure InvivoGen Cat# tlrl-pelps Filgotinib TargetMol Cat# T1929 Stattic MedChem Express Cat# HY-13818 Fludarabine MedChem Express Cat# HY-B0069 Ruxolitinib Cayman Chemical Cat# 11609-5 Tofacitinib MedChem Express Cat# HY-40354 OVA323–399 Bachem Cat# H-5532 CD11c MACS Micro Bead System Miltenyi Biotec Cat# 130-125-835 Naive CD4+ T Cell Isolation Kit, mouse Miltenyi Biotec Cat# 130-104-453 CFSE Thermo Fisher Scientific Cat# 65-0850-84 Recombinant Mouse TGF-beta 1 R&D Cat# 7666-MB-005/CF Animal-Free Recombinant Human IL-2 Peprotech Cat# AF-200-02 Recombinant mouse PD-L1 Fc-Fusion protein Kerafast Cat# Pr00112-1.9 Recombinant human IgG1 Fc control protein R&D Cat# 110-HG-100 Adult brain dissociation kit Miltenyi Biotec Cat# 130-107-677 Fixable Viability Dye eFluorTM 780 Thermo Fisher Scientific Cat# 65-0865-14 SYTOXTM AADvancedTM Thermo Fisher Scientific Cat# S10274 FoxP3/Transcription factor staining buffer set Thermo Fisher Scientific Cat# 00-5523-00 eBioscienceTM Intracellular fixation & permeabilization buffer set Thermo Fisher Scientific Cat# 88-8824-00 True-Phos Buffer Set BioLegend Cat# 425401 Ionomycin Sigma Aldrich Cat# I0634 PMA Sigma Aldrich Cat# P1585 Brefeldin A solution Thermo Fisher Scientific Cat# 00-4506-51 High-Capacity cDNA Reverse Transcription Kit Thermo Fisher Scientific Cat# 4368814 TaqManTM Gene Expression Master Mix Thermo Fisher Scientific Cat# 4369016 Luna Master Mix New England BioLabs Cat# M3003E Critical commercial assays LEGENDplexTM Mouse Inflammation Panel BioLegend Cat# 740446 LEGENDplexTM Mouse Th Panel BioLegend Cat# 740741 Experimental models: Organisms/strains Mouse: Jak1tm1a(EUCOMM)Hmgu EUMMCR MGI:4456720 Mouse: ROSA26::FLPe knock in; B6.129S4-Gt(ROSA)26Sortm1(FLP1)Dym/RainJ The Jackson Laboratory Cat# JAX:009086, RRID:IMSR_JAX:009086 Mouse: CD11c cre; B6.Cg-Tg(Itgax-cre)11Reiz/J mice The Jackson Laboratory Cat# JAX:008068, RRID:IMSR_JAX:008068 (Continued on next page) Cell Reports 38, 110420, February 22, 2022 e3
Article Title: Succinate uptake by T cells suppresses their effector function via inhibition of mitochondrial glucose oxidation.
Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies anti-CD69-APC Biolegend Clone FN50, Cat# 310910; RRID:AB_314845 anti-CD25-BV605 Biolegend Clone BC96, Cat# 302632; RRID:AB_11218989 anti-CD107a-FITC BD Pharmingen Clone H4A3, Cat# 560949; RRID:AB_2033934 anti-IFN-g-FITC Biolegend Clone B27, Cat# 506504; RRID:AB_315437 anti-TNF-a-PE Biolegend Clone Mab11, Cat# 502909; RRID:AB_315261 anti-IFN-g capture (ELISA) Bio-Rad Clone AbD00676, Cat# HCA043; RRID:AB_906045 anti-IFN-g biotinylated (ELISA) Bio-Rad Clone 2503 Cat# HCA044P; RRID:AB_10614712 anti-HIF-1a [H1alpha67] (WB) Abcam Cat# ab1; RRID:AB_296474 anti-SUCNR1 (WB) Novus Biologicals Cat# NBP1-00861; RRID:AB_1503315 anti-CD3 (mouse) Biolegend Clone 145-2C11 Cat# 100340; RRID:AB_11149115 anti-CD28 (mouse) Biolegend Clone 37.51 Cat# 102116; RRID:AB_11147170 anti-IFN-g-FITC (mouse) Biolegend Clone XMG1.2, Cat# 505806; RRID:AB_315400 anti-TNF-a-PE (mouse) Biolegend Clone MP6-XT22, Cat# 506305; RRID:AB_315426 Chemicals, peptides, and recombinant proteins RPMI1640 Cell Culture Medium Sigma Aldrich Cat# F9665 Penicillin/Streptomycin Thermo Fisher Cat# 15140122 Recombinant human IL-2 PeproTech Cat# 200-02 Immunocult Human CD3/CD28 T cell activator Stemcell Technologies Cat# 10991 Sodium Succinate Sigma Aldrich Cat# 14160 AZD-3965 Selleckchem Cat# S7339 b-chloro-L-alanine Sigma Aldrich Cat# C9033 Sodium Pyruvate Sigma Aldrich Cat# P8574 Dichloroacetate Sigma Aldrich Cat# 347795 Sodium Metavanadate Sigma Aldrich Cat# 72060 Cell Activation Cocktail Biolegend Cat# 423304 Fixation/permeabilization solutions eBioscience Cat# 00-5523-00 CellTrace Violet Thermo Fisher Cat# C34557 Recombinant IFN-g standard (ELISA) Bio-Rad Cat# PHP050 streptavidin-HRP Sigma Aldrich Cat# E2866 TMB substrate Bio-Rad Cat# BUF062B MitoSpy Orange Biolegend Cat# 424804 Laemmli buffer Sigma Aldrich Cat# 38733 RIPA buffer Thermo Fisher Scientific Cat# 89900 Clarity Western ECL Substrate Bio-Rad Cat# 170-5061 U-13C6 glucose CK Isotopes Cat# CLM-1396-1 13C4 sodium succinate Sigma Aldrich Cat# 491985 Methoxyamine hydrochloride in pyridine Thermo Fisher Scientific Cat# 25104 Lipofectamine RNAiMAX Thermo Fisher Cat# 13778075 Fixable Viability Dye eFluorTM 780 Thermo Fisher Scientific Cat# 65-0865-14 4C (SUCNR1 antagonist) Provided by Prof S Pluchino N/A (Continued on next page) Cell Reports 40, 111193, August 16, 2022 e1
Knock-In:Article Title: JAK1 signaling in dendritic cells promotes peripheral tolerance in autoimmunity through PD-L1-mediated regulatory T cell induction.
Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Chemicals, peptides, and recombinant proteins Myelin oligodendrocyte glycoprotein (MOG)35–55 Charitè Berlin NA Incomplete Freund’s adjuvant Sigma-Aldrich Cat# F5506 Mycobacterium tuberculosis H37Ra BD Cat# 231141 Pertussis toxin List Biological Laboratories Cat# 181 recombinant murine GM-CSF R&D Cat# 415-ML-050 recombinant murine IL-4 Miltenyi Biotec Cat# 130-097-761 recombinant murine IFN-ɣ Miltenyi Biotec Cat# 130-105-774 recombinant murine IL-10 R&D Cat# 417-ML-005 Lipopolysaccharide ultra pure InvivoGen Cat# tlrl-pelps Filgotinib TargetMol Cat# T1929 Stattic MedChem Express Cat# HY-13818 Fludarabine MedChem Express Cat# HY-B0069 Ruxolitinib Cayman Chemical Cat# 11609-5 Tofacitinib MedChem Express Cat# HY-40354 OVA323–399 Bachem Cat# H-5532 CD11c MACS Micro Bead System Miltenyi Biotec Cat# 130-125-835 Naive CD4+ T Cell Isolation Kit, mouse Miltenyi Biotec Cat# 130-104-453 CFSE Thermo Fisher Scientific Cat# 65-0850-84 Recombinant Mouse TGF-beta 1 R&D Cat# 7666-MB-005/CF Animal-Free Recombinant Human IL-2 Peprotech Cat# AF-200-02 Recombinant mouse PD-L1 Fc-Fusion protein Kerafast Cat# Pr00112-1.9 Recombinant human IgG1 Fc control protein R&D Cat# 110-HG-100 Adult brain dissociation kit Miltenyi Biotec Cat# 130-107-677 Fixable Viability Dye eFluorTM 780 Thermo Fisher Scientific Cat# 65-0865-14 SYTOXTM AADvancedTM Thermo Fisher Scientific Cat# S10274 FoxP3/Transcription factor staining buffer set Thermo Fisher Scientific Cat# 00-5523-00 eBioscienceTM Intracellular fixation & permeabilization buffer set Thermo Fisher Scientific Cat# 88-8824-00 True-Phos Buffer Set BioLegend Cat# 425401 Ionomycin Sigma Aldrich Cat# I0634 PMA Sigma Aldrich Cat# P1585 Brefeldin A solution Thermo Fisher Scientific Cat# 00-4506-51 High-Capacity cDNA Reverse Transcription Kit Thermo Fisher Scientific Cat# 4368814 TaqManTM Gene Expression Master Mix Thermo Fisher Scientific Cat# 4369016 Luna Master Mix New England BioLabs Cat# M3003E Critical commercial assays LEGENDplexTM Mouse Inflammation Panel BioLegend Cat# 740446 LEGENDplexTM Mouse Th Panel BioLegend Cat# 740741 Experimental models: Organisms/strains Mouse: Jak1tm1a(EUCOMM)Hmgu EUMMCR MGI:4456720 Mouse: ROSA26::FLPe knock in; B6.129S4-Gt(ROSA)26Sortm1(FLP1)Dym/RainJ The Jackson Laboratory Cat# JAX:009086, RRID:IMSR_JAX:009086 Mouse: CD11c cre; B6.Cg-Tg(Itgax-cre)11Reiz/J mice The Jackson Laboratory Cat# JAX:008068, RRID:IMSR_JAX:008068 (Continued on next page) Cell Reports 38, 110420, February 22, 2022 e3
Article Title: Succinate uptake by T cells suppresses their effector function via inhibition of mitochondrial glucose oxidation.
Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies anti-CD69-APC Biolegend Clone FN50, Cat# 310910; RRID:AB_314845 anti-CD25-BV605 Biolegend Clone BC96, Cat# 302632; RRID:AB_11218989 anti-CD107a-FITC BD Pharmingen Clone H4A3, Cat# 560949; RRID:AB_2033934 anti-IFN-g-FITC Biolegend Clone B27, Cat# 506504; RRID:AB_315437 anti-TNF-a-PE Biolegend Clone Mab11, Cat# 502909; RRID:AB_315261 anti-IFN-g capture (ELISA) Bio-Rad Clone AbD00676, Cat# HCA043; RRID:AB_906045 anti-IFN-g biotinylated (ELISA) Bio-Rad Clone 2503 Cat# HCA044P; RRID:AB_10614712 anti-HIF-1a [H1alpha67] (WB) Abcam Cat# ab1; RRID:AB_296474 anti-SUCNR1 (WB) Novus Biologicals Cat# NBP1-00861; RRID:AB_1503315 anti-CD3 (mouse) Biolegend Clone 145-2C11 Cat# 100340; RRID:AB_11149115 anti-CD28 (mouse) Biolegend Clone 37.51 Cat# 102116; RRID:AB_11147170 anti-IFN-g-FITC (mouse) Biolegend Clone XMG1.2, Cat# 505806; RRID:AB_315400 anti-TNF-a-PE (mouse) Biolegend Clone MP6-XT22, Cat# 506305; RRID:AB_315426 Chemicals, peptides, and recombinant proteins RPMI1640 Cell Culture Medium Sigma Aldrich Cat# F9665 Penicillin/Streptomycin Thermo Fisher Cat# 15140122 Recombinant human IL-2 PeproTech Cat# 200-02 Immunocult Human CD3/CD28 T cell activator Stemcell Technologies Cat# 10991 Sodium Succinate Sigma Aldrich Cat# 14160 AZD-3965 Selleckchem Cat# S7339 b-chloro-L-alanine Sigma Aldrich Cat# C9033 Sodium Pyruvate Sigma Aldrich Cat# P8574 Dichloroacetate Sigma Aldrich Cat# 347795 Sodium Metavanadate Sigma Aldrich Cat# 72060 Cell Activation Cocktail Biolegend Cat# 423304 Fixation/permeabilization solutions eBioscience Cat# 00-5523-00 CellTrace Violet Thermo Fisher Cat# C34557 Recombinant IFN-g standard (ELISA) Bio-Rad Cat# PHP050 streptavidin-HRP Sigma Aldrich Cat# E2866 TMB substrate Bio-Rad Cat# BUF062B MitoSpy Orange Biolegend Cat# 424804 Laemmli buffer Sigma Aldrich Cat# 38733 RIPA buffer Thermo Fisher Scientific Cat# 89900 Clarity Western ECL Substrate Bio-Rad Cat# 170-5061 U-13C6 glucose CK Isotopes Cat# CLM-1396-1 13C4 sodium succinate Sigma Aldrich Cat# 491985 Methoxyamine hydrochloride in pyridine Thermo Fisher Scientific Cat# 25104 Lipofectamine RNAiMAX Thermo Fisher Cat# 13778075 Fixable Viability Dye eFluorTM 780 Thermo Fisher Scientific Cat# 65-0865-14 4C (SUCNR1 antagonist) Provided by Prof S Pluchino N/A (Continued on next page) Cell Reports 40, 111193, August 16, 2022 e1
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