KEY RESOURCES TABLE
| REAGENT or RESOURCE | SOURCE | IDENTIFIER |
|---|---|---|
| Antibodies | ||
| CD4-PE Clone GK1.5 (1:200) | Biolegend | #100408 |
| CD25-PE Clone PC61 (1:200) | Biolegend | #102008 |
| IRF4-PE Clone IRF4.3F4 (1:200) | Biolegend | #646404 |
| F4/80-PE Clone BM8 (1:200) | Biolegend | #123110 |
| CD185-PE Clone L138D7 (1:200) | Biolegend | #145504 |
| CD90.1-PE Clone HIS51 (1:200) | Thermo Fisher | #12-0900-81 |
| CD45RB-PE Clone C363-16A (1:200) | Biolegend | #103308 |
| Ki-67-PE Clone 16A8 (1:200) | Biolegend | #652404 |
| TGF-β1,-β2,-β3-PE Clone 1D11 (1:200) | R&D Systems | #IC1835P |
| Tbet-PE/Cy7 Clone 4B10 (1:200) | Biolegend | #644824 |
| F4/80-PE/Cy7 Clone BM8 (1:200) | Biolegend | #123114 |
| Bcl6-PE/Cy7 Clone 7D1 (1:200) | Biolegend | #358512 |
| Helios-PE/Cy7 Clone 22F6 (1:200) | Biolegend | #137236 |
| pSTAT5-PE/Cy7 Clone SRBCZY (1:200) | Thermo Fisher | #25-9010-42 |
| CD8a-APC Clone 53-6.7 (1:200) | Biolegend | #100712 |
| IFN-γ-APC Clone XMG1.2 (1:200) | Biolegend | #505810 |
| Foxp3-APC Clone FJK-16s (1:200) | Thermo Fisher | #17-5773-82 |
| pS6-APC Clone cupk43k (1:200) | Therno Fisher | #17-9007-42 |
| GL7-APC Clone GL7 (1:200) | Biolegend | #144606 |
| I-A/I-E-APC Clone M5/114.15.2 (1:200) | Biolegend | #107614 |
| CD98-APC Clone RL388 (1:200) | Biolegend | #128210 |
| pS473-APC Clone M89-61 (1:200) | BD Biosciences | #560343 |
| Granzyme B-APC Clone NGZB (1:200) | Thermo Fisher | #50-8898-62 |
| IgG (H+L)-APC (1:500) | Thermo Fisher | #A21244 |
| CD3ε-APC-Cy7 Clone 145-2C11 (1:200) | Biolegend | #100330 |
| CD86-FITC Clone GL-1 (1:200) | Biolegend | #105006 |
| Perforin-FITC Clone eBioOMAK-D (1:200) | Thermo Fisher | #11-9392-82 |
| RORγT-BV421 Clone Q31-378 (1:200) | BD Biosciences | #562894 |
| CD95-BV421 Clone Jo2 (1:200) | BD Biosciences | #562633 |
| CD152-BV421 Clone UC10-4B9 (1:200) | Biolegend | #106312 |
| CD134-BV421 Clone OX-86 (1:200) | Biolegend | #119411 |
| CD62L-Pacific Blue Clone MEL-14 (1:200) | Biolegend | #104424 |
| CD25-Pacific Blue Clone PC61 (1:200) | Biolegend | #102022 |
| TNF-α-Pacific Blue Clone MP6-XT22 (1:200) | Biolegend | #506318 |
| pmTOR-Pacific Blue Clone MRRBY (1:200) | Biolegend | #48-9718-42 |
| c-Myc-Pacific Blue Clone D84C12 (1:200) | Cell Signalling | #14426S |
| Foxp3-V450 Clone MF23 (1:200) | BD Biosciences | #561293 |
| CD44-PerCP/Cy5.5 Clone IM7 (1:200) | Biolegend | #103032 |
| CD86-PerCP Clone GL-1 (1:200) | Biolegend | #105026 |
| CD45R/B220-BV510 Clone RA3-6B2 (1:200) | Biolegend | #103248 |
| CD69-BV605 Clone H1.2F3 (1:200) | Biolegend | #104530 |
| IL-17A-BV605 Clone TC11-18H10 (1:200) | BD Biosciences | #564169 |
| CD279-BV605 Clone 29F.1A12 (1:200) | Biolegend | #135220 |
| CD4-BV785 Clone GK1.5 (1:200) | Biolegend | #100453 |
| CD19-BV785 Clone 6D5 (1:200) | Biolegend | #115543 |
| Glut1-Alexa Fluor®488 Clone EPR3915 (1:200) | Abcam | #ab195359 |
| pSMAD3 Ser423, Ser425 | Thermo Fisher | #44-246G |
| pAMPK alpha-1,2 (Thr172) | Thermo Fisher | #44-1150G |
| Purified anti-mouse CD3ε Clone 145-2C11 | Biolegend | #100340 |
| Purified anti-mouse CD28 Clone | Biolegend | #102112 |
| Human CD3/CD28 T Cell activator | Stemcell Technologies | #10971 |
| Anti-Mouse IFN-γ Clone XMG1.2 | BD Biosciences | #554408 |
| Biological Samples | ||
| Healthy control buffy coat | Croix-Rouge Luxembourgeoise | N/A |
| Chemicals, Peptides, and Recombinant Proteins | ||
| RPMI 1640 (without L-Glutamine) | Lonza | #BE12-167F |
| RMPI 1640 (without glucose, serine, glycine) | Teknova | #50-190-8105 |
| DMEM (with glucose and L-Glutamine) | Lonza | #BE12-604F |
| Hank’s Balanced Salt Solutions | Lonza | #BE10-543F |
| SILAC RPMI 1640 Flex Media | Gibco | #15347143 |
| Seahorse XF base medium without phenol red | Agilent Technologies | #103335-100 |
| PBS | Lonza | #BE17-516F |
| FBS | Sigma-Aldrich | #TMS-013-B |
| Penicillin/Streptomycin | Gibco | #11548876 |
| L-Glutamine | Sigma-Aldrich | #G3126-100G |
| 2-mercaptoethanol | Gibco | #11508916 |
| Sodium pyruvate | Gibco | #12539059 |
| Recombinant human TGF-β | R&D Systems | #240-B-002 |
| recombinant human IL-2 | Miltenyi Biotec | #130-120-333 |
| 2-Deoxy-D-glucose | Sigma-Aldrich | #D6134-1G |
| D-(+)-glucose | Sigma-Aldrich | #G8270-5KG |
| Oligomycin A | Sigma-Aldrich | #75351-5MG |
| FCCP | Sigma-Aldrich | #C2920-10MG |
| Antimycin A | Sigma-Aldrich | #A8674-25MG |
| Rotenone | Sigma-Aldrich | #R8875-1G |
| Phorbol 12-myristate 13-acetate (PMA) | Sigma-Aldrich | #P8139-1MG |
| Calcium Ionophore | Sigma-Aldrich | #C7522-1MG |
| N-Acetyl-L-cysteine | Sigma-Aldrich | #A7250-50G |
| L-Glutathione reduced | Sigma-Aldrich | #G4251-10G |
| Rapamycin | Invivogen | #tlrl-rap |
| Rapamycin | LC Laboratories | N/A |
| L-(+)-α-Phenylglycine | Sigma-Aldrich | #237647-25G |
| 4-Hydroxytamoxifen | Sigma-Aldrich | #H6278-10MG |
| PKUMDL-WQ-2101 | Sigma-Aldrich | #SML1970-5MG |
| L-buthionine-sulfoximine | Sigma-Aldrich | #B2515-1G |
| SHIN1 | Aobious | #AOB36697 |
| Thymidine, [6-3H]-, 5mCi (185MBq) | Perkin Elmer | # NET355005MC |
| SYBR™ Fast Green Master Mix | Applied Biosystems | # 10459604 |
| Corning™ Cell-Tak Cell and Tissue Adhesive | Thermo Fisher | # 10317081 |
| 13C6-glucose | Cambridge Isotope Lab | # CLM-1396 |
| 13C5-L-Glutamine | Cambridge Isotope Lab | # CLM-1822-H |
| 13C3-serine | Cambridge Isotope Lab | #CLM-1574-H |
| GolgiPlug™ | BD Biosciences | #555029 |
| Cell Trace Violet | Thermo Fisher | #C34557 |
| Mitotracker Deep Red | Thermo Fisher | # M22426 |
| Molecular Probes™ Carboxy-H2DCFDA | Thermo Fisher | #11500146 |
| Molecular Probes™ 2-NBDG | Thermo Fisher | #11569116 |
| Zombie NIR™ Fixable Viability Kit | Biolegend | #423106 |
| Saponin | Sigma-Aldrich | #S4521-25G |
| Formaldehyde | Sigma-Aldrich | #252549-1L |
| Hepes | Sigma-Aldrich | #H4034-100G |
| EDTA | Sigma-Aldrich | #E9884-100G |
| Polyethylene glycol | Sigma-Aldrich | #P1458-50ML |
| Ethanol | VWR | #20821330 |
| Tween 80 | Sigma-Aldrich | #P1754-1L |
| Methanol | Sigma-Aldrich | #1060351000 |
| Chloroform | Sigma-Aldrich | #34854-1L-M |
| Trichloroacetic acid | Sigma-Aldrich | #T6399-100G |
| DNAse I | Sigma-Aldrich | #10104159001 |
| Liberase | Sigma-Aldrich | #5401020001 |
| Percoll® | Sigma-Aldrich | #GE17-0891-01 |
| HiSep™ LSM 1077 | HiMedia Laboratory | #LS001-500ML |
| Critical Commercial Assays | ||
| Naive CD4+ T Cell Isolation Kit | Miltenyi Biotec | #130-104-453 |
| CD4+ T Cell Isolation Kit | Miltenyi Biotec | #130-104-454 |
| CD90.1 Microbeads | Miltenyi Biotec | #130-094-523 |
| CD4+ CD25+ Regulatory T Cell Isolation Kit | Miltenyi Biotec | #130-091-041 |
| Tumor Dissociation Kit | Miltenyi Biotec | #130-096-730 |
| CD45 (TIL) Microbeads | Miltenyi Biotec | #130-110-618 |
| Naive CD4+ T Cell Isolation Kit II, human | Miltenyi Biotect | #130-094131 |
| Seahorse XFe96 Fluxpak | Agilent Technologies | #102416-100 |
| Foxp3/Transcription Factor Staining Buffer Set | Thermo Fisher | #15151976 |
| Fixation/Permeabilization solution kit | BD Biosciences | #554701 |
| Lyse/Fix Buffer 5x | BD Biosciences | #558049 |
| Perm buffer III | BD Biosciences | #558050 |
| QuantiTect Reverse Transcription Kit | Qiagen | #205314 |
| RNA 6000 Nano Kit | Agilent Technologies | #5067-1511 |
| NucleoSpin RNA 250 | Macherey-Nagel | #740 955 250 |
| IFN gamma Mouse Uncoated ELISA Kit | Thermo Fisher | #88-7314-88 |
| TNF alpha Mouse Uncoated ELISA Kit | Thermo Fisher | #88-7324-88 |
| Invitrogen ™ IL-2 Mouse Uncoated ELISA Kit | Thermo Fisher | #15520997 |
| Experimental Models: Cell Lines | ||
| B16F10 | Dr. Philipp Lang | |
| MC38 | ATCC | |
| Experimental Models: Organisms/Strains | ||
| Gclcfl/fl : B6 | Maketal., 2017 | N/A |
| Foxp3YFP-Cre : B6 | The Jackson laboratory | #016959 |
| Foxp3eGFP-Cre-ERT2 : B6 | The Jackson laboratory | #016961 |
| Rag1−/− : B6 | The Jackson laboratory | #002216 |
| IFNγ−/− : B6 | The Jackson laboratory | #002287 |
| Deposited Data | ||
| RNA-Seq | This paper | |
| Oligonucleotides | ||
| Gclc: F: G G CTCTCTG CACCATCACTT R :GTTAGAGTACCGAAGCGGGG |
This paper | |
| Foxp3: F:CCCATCCCCAGGAGTCTTG R : ACCAT G ACT AG GG G CACT GTA |
This paper | |
| Slc1a4: F:TGCTCTGGCGTTCATCATCA R :AGTGAATG CGG CAACCACAA |
This paper | |
| Slc1a5: F :TGG CCAG CAAG ATTGTG GAG AT R :TTTGCGGGTGAAGAGGAAGT |
This paper | |
| HPRT: F:TCAGTCAACGGGGGACATAAA R :GGGGCTGTACTGCTTAACCAG |
This paper | |
| TBP: F :G AAG AAC AAT CCAG ACTAG CAG CA R : CCTTATAG G G AACTT C AC AT C AC AG |
This paper | |
| PHGDH: F: ATG GCCTTCG CAAATCTG C R :AGTTCAGCTATCAGCTCCTCC |
This paper | |
| sgRNA Phgdh C*U*U*GCCUUGCCUUGCCCAUG G*G*C*AAGAGCUCACCUUCUUC U*U*C*UUACAGGCAGAUUCCCC |
Synthego | N/A |
| sgRNA Slc1a4 A*C*C*AGGCUGCAAACCACCAG G*C*A*UCUCGCCCGGGAAGGCC C*C*U*CAGCGCCGCGCCCAUGC |
Synthego | N/A |
| Recombinant DNA | ||
| pMIT-Foxp3-CD90.1 | Dr. Michael Lohoff | |
| pMIT-CD90.1 | Dr. Michael Lohoff | |
| pMIG-RI-STAT5-CA | Dr. Michael Lohoff | |
| pMIG-RI | Dr. Russell Jones | |
| pMIG-RI-Gclc | This paper | |
| pMIG-RI-Smad3 | This paper | |
| Software and Algorithms | ||
| FlowJo Software | Tree Star | N/A |
| Continued | ||
| Graphpad Prism | GraphPad Software, Inc | N/A |
| Wave Software | Agilent | N/A |
| Adobe Illustrator | Adobe systems | N/A |