KEY RESOURCES TABLE
| REAGENT or RESOURCE | SOURCE | IDENTIFIER |
|---|---|---|
| Antibodies | ||
| Anti-mouse CD3 | BioXCell | Clone: 145–2C11; Cat# BE0001–1; RRID: AB_1107634 |
| Anti-mouse CD28 | BioXCell | Clone: 37.51; Cat# BE0015–1; RRID: AB_1107624 |
| Anti-mouse IL4 | BioXCell | Clone: 11B11; Cat# BE0045; RRID: AB_1107707 |
| Anti-mouse IFNγ | BioXCell | Clone: XMG1.2; Cat# BE0055; RRID: AB_1107694 |
| CD4-PE | eBioscience | Clone: GK5.1; Cat# 12–0041-83; RRID: AB_465506 |
| RORγ-APC | eBioscience | Clone: AFKJS-9; Cat# 17–6988-80; RRID: AB_10609207 |
| FOXP3 -PE | eBioscience | Clone: FJK-16 s; Cat# 12–5773-82; RRID: RRID:AB_465936 |
| IRF-4-eFluor 660 | eBioscience | Clone: 3E4; Cat# 50–9858-80; RRID: AB_2574393 |
| CXCR3-PE | eBioscience | Clone: CXCR3–173; Cat# 12–1831-80; RRID: AB_1210734 |
| CD45.2-PE | eBioscience | Clone: 104; Cat# 12–0454-81; RRID: AB_465678 |
| CD90.1-APC | eBioscience | Clone: HIS51; Cat# 17–0900-82; RRID: AB_1272252 |
| CD90.1-Pacific Blue | eBioscience | Clone: HIS51; Cat# 48–0900-80; RRID: AB_1272254 |
| CD4-PE/Cy7 | Biolegend | Clone: GK5.1; Cat# 100422; RRID: AB_312707 |
| CD4-APC | Biolegend | Clone: GK5.1; Cat# 100412; RRID: AB_312697 |
| CD4-APC/Cy7 | Biolegend | Clone: GK5.1; Cat# 100414; RRID: AB_312699 |
| CD8-PE/Cy7 | Biolegend | Clone: 53–6.7; Cat# 100722; RRID: AB_312761 |
| CD8-APC | Biolegend | Clone: 53–6.7; Cat# 100712; RRID: AB_312751 |
| CD8-FITC | Biolegend | Clone: 53–6.7; Cat# 100706; RRID: AB_312745 |
| IL17-PE | Biolegend | Clone: TC11–18H10.1; Cat# 506904; RRID: AB_315464 |
| IL17-Pacific Blue | Biolegend | Clone: TC11–18H10.1; Cat# 506918; RRID: AB_893544 |
| IFNγ-PE | Biolegend | Clone: XMG1.2; Cat# 505808; RRID: AB_315402 |
| IFNγ-Alexa647 | Biolegend | Clone: XMG1.2; Cat# 505814; RRID: AB_493314 |
| CCR7-PE | Biolegend | Clone: 4B12; Cat# 120105; RRID: AB_389357 |
| CD36-APC | Biolegend | Clone: HM36; Cat# 102612; RRID:AB_2072639 |
| CD62L-APC | Biolegend | Clone: MEL14; Cat# 104412; RRID AB_313099 |
| CD44-PerCP/cye5.5 | Biolegend | Clone: IM7; Cat# 103032; RRID AB_2076204 |
| CD45.1-APC | BD Biosciences | Clone: A20; Cat# 558701; RRID: AB_1645214 |
| Vβ13-FITC | BD Biosciences | Clone: MR1 2–3; Cat# 553204; RRID: AB_394706 |
| KLRG1-PerCP/Cye5.5 | BD Biosciences | Clone: 2F1; Cat# 563595 |
| Mouse S1P1 /EDG-1 PE | R&D System | Clone: # 713412; Catalog# FAB7089P |
| Mouse TGF-beta RII FlTC | R&D System | Clone: # Ile24Asp184 Accession # Q62312; Catalog# FAB532F |
| Mouse anti-Akt (pS473) PE | BD Biosciences | Clone # M89–61 Cat# 560378 RRID:AB_1645328 |
| Mouse anti-Akt (pS473) | Cell Signaling Technology | Clone: N/A Cat# 9271 RRID N/A |
| Anti-Goat Alexa647 | Thermo Fisher Scientific | Clone: N/A; Cat# A21447; RRID: AB_141844 |
| pFoxo1 (S256) | Cell Signaling Technology | Clone: N/A; Cat# 9461; RRID: AB_329831 |
| Phospho-S6 Ribosomal Protein (Ser235/236)-Alexa647 | Cell Signaling Technology | Clone: D57.2.2E; Cat# 4851; RRID: AB_916160 |
| Foxo1 | Cell Signaling Technology | Clone: C29H4; Cat# 2880S; RRID: AB_2106495 |
| Anti-Rabbit HRP | Cell Signaling Technology | Clone: N/A; Cat# 7074S; RRID: N/A |
| Anti-Rabbit PE | Jackson ImmunoResearch Laboratories | Clone: N/A; Cat# 111–116-144; RRID: AB_2337985 |
| Anti-Rabbit Alexa647 | Jackson ImmunoResearch Laboratories | Clone: N/A; Cat# 111–607-003; RRID: AB_2338084 |
| Anti-mouse PPARγ Alexa647 | Santa Cruz Biotechnology | Clone: E-8; Cat# Sc7273; |
| PPARγ (C26H12) Rabbit mAb | Cell Signaling Technology | Clone C26H12, Cat# 2435 |
| β-Tubulin Ab | Cell Signaling Technology | Clone: N/A; Cat#2146S |
| TATA-binding protein (TBP) Ab | Cell Signaling Technology | Clone: N/A; Cat#8515S |
| inVivoMAb anti-mouse PD-1 | BioxCell | Cat# BE0146 Clone RMPI 14 RRID: RRID:AB_10949053 |
|
Chemicals, Peptides, and Recombinant Proteins | ||
| GW9662 | Cayman Chemical | Cat# 70785 |
| TZD | Cayman Chemical | Cat# 71740 |
| Orlistat | Cayman Chemical | Cat# 10005426 |
| Etomoxir | Cayman Chemical | Cat# 11969 |
| AS1842856 | Sigma Aldrich | Cat# 344355 |
| AKTi | Cayman Chemical | Cat# 14870 |
| SC-79 | Sigma Aldrich | Cat# SML0749 |
| FTY720 | Sigma Aldrich | Cat# SML0700 |
| PF-543 | Calbiochem | Cat# 567741 |
| W146 | Cayman Chemical | Cat# 10009109 |
| JTE-013 | Cayman Chemical | Cat# 10009458 |
| S1P | Avanti Polar Lipids | Cat# 860662 |
| Oleic Acid | Sigma | Cat# O3008 |
| BODIPY 493/503 | Thermo Fisher Scientific | Cat# D3922 |
| BODIPY 558/568 C12 | Thermo Fisher Scientific | Cat# D38835 |
| 2-Deoxy-D-glucose (2DG) | Sigma Aldrich | Cat# D6134 |
| Antimycin A | Sigma Aldrich | Cat# A8674 |
| Rotenone | Sigma Aldrich | Cat# R8875 |
| Oligomycin | Sigma Aldrich | Cat# O4876 |
| FCCP | Sigma Aldrich | Cat# C2920 |
| IMDM | GE Healthcare, HyClone | Cat# SH30228.01 |
| RPMI-1640 (Glucose free) | Thermo Fisher Scientific | Cat# 11879–020 |
| Penicillin-Streptomycin | Corning | Cat# 30–001-CI |
| Fetal Bovine Serum (FBS) | Atlanta Biologicals | Cat# S11150 |
| rIL12 | Biolegend | Cat# 577004 |
| rIL6 | Biolegend | Cat# 575704 |
| rTGFβ | Biolegend | Cat# 580702 |
| rhIL2 | NCI, Biological Resources Branch | https://ncifrederick.cancer.gov/research/brb/productDataSheets/cytokineHumanInterleukins/IL-2Bulk.aspx |
| Foxp3 / Transcription Factor Staining Buffer Set | Thermo Fisher Scientific | Cat# 00–5523 |
| Fixation/Permeabilization Solution Kit | BD Biosciences | Cat# 554714 |
| hgp10025–33 peptide (KVPRNQDW) | Genscript | Cat# RP20344 |
| Nucleofector Kits for Mouse T Cells | Lonza | Cat# VPA-1006 |
| RIPA Lysis Buffer | Thermo Fisher Scientific | Cat# 89900 |
| NE-PER Nuclear and Cytoplasmic Extraction Reagents | Thermo Fisher Scientific | Cat# 78833 |
| rIL12 | Biolegend | Cat# 577004 |
|
Critical Commercial Assays | ||
| CellTrace CFSE Cell Proliferation Kit | Thermo Fisher Scientific | Cat# C34554 |
| TransAM® FKHR (FOXO1) Transcription Factor ELISA Kits | Active Motif | Cat# 46396 |
| PPAR 96-Well Strip Plate | Cayman Chemical | Cat#10006887 |
| Triglycerides Reagent | ThermoFisher Scientific | Cat# TR22421 |
| Total Cholesterol Reagents | ThermoFisher Scientific | Cat# TR13421 |
| Adenosine 5′-triphosphate (ATP) Bioluminescent Assay Kit | Sigma | Cat# FLAA-1KT |
| iScript cDNA Synthesis Kit | Biorad | Cat# 1708891 |
| SsoAdvanced Universal SYBR® Green Supermix | Biorad | Cat# 1725274 |
|
Deposited Data | ||
| RNA-seq | This Paper | Accession number GSE129489 |
|
Mouse qPCR primers: Oligonucleotide Sequences (5′−3′) | ||
| Cpt1a Forward TCTATGAGGGCTCGCG | This Paper | N/A |
| Cpt1a Reverse CGTCAGGGTTGTAGCA | This Paper | N/A |
| IL23r Forward TTCAGATGGGCATGAATGTTTCT | This Paper | N/A |
| IL23r Reverse CCAAATCCGAGCTGTTGTTCTAT | This Paper | N/A |
| Csf2 Forward GGCCTTGGAAGCATGTAGAGG | This Paper | N/A |
| Csf2 Reverse GGAGAACTCGTTAGAGACGACTT | This Paper | N/A |
| IL22 Forward ATGAGTTTTTCCCTTATGGGGAC | This Paper | N/A |
| IL22 Reverse GCTGGAAGTTGGACACCTCAA | This Paper | N/A |
| IL9 Forward CATCAGTGTCTCTCCGTCCCAA CTGAT | This Paper | N/A |
| IL9 Reverse GATTTCTGTGTGGCATTGGTCAG | This Paper | N/A |
| Lef1 Forward CTCGTCGCTGTAGGTGATGA | This Paper | N/A |
| Lef1 Reverse AAATGGGTCCCTTTCTCCAC | This Paper | N/A |
| Nanog Forward GCAAGAATAGTTCTCGG GATGAA | This Paper | N/A |
| Nanog Reverse TCTTCCTGGTCCCCACAGTTT | This Paper | N/A |
| Sphk1 Forward ATGGAACCAGTAGAATGCCCT | This Paper | N/A |
| Sphk1 Reverse TCCGTTCGGTGAGTAT CAGTTTA | This Paper | N/A |
| Tcf7 Forward GTGGACTGCTGAAATGTTCG | This Paper | N/A |
| Tcf7 Reverse AGCATCCGCAGCCTCAAC | This Paper | N/A |
| β-actin Forward ACGTAGCCATCCAGG CTGGTG | This Paper | N/A |
| β-actin Reverse TGGCGTGAGGGAGAGCAT | This Paper | N/A |
|
Experimental Models: Cell Lines | ||
| B16-F10 | ATCC | CRL-6475 |
| B16-F10-A2+ | Rolf Kiessling, Karolinska Institute, Stockholm, Sweden. Mycoplasma testing was done in Mehrotra lab. | N/A |
|
Experimental Models: Organisms/Strains | ||
| C57BL/6 | Jackson Laboratory | Stock# 000664 |
| C57BL/6-Tg(HLA-A2.1)1Enge/J | Jackson Laboratory | Stock# 003475 |
| B6.129S7-Rag1tm1Mom/J | Jackson Laboratory | Stock# 002216 |
| B6.Cg-Thy1a/Cy Tg(TcraTcrb)8Rest/J | Jackson Laboratory | Stock# 005023 |
| PPARγfl/flCD4cre | Robert B Clark, University of Connecticut Health Center | Wohlfert et al., 2007 |
|
Software and Algorithms | ||
| FlowJo 10.2 | TreeStar, OR | https://www.flowjo.com/solutions/flowjo/downloads/ |
| Prism 8 | GraphPad | https://www.graphpad.com/scientific-software/prism/ |
| Agilent Seahorse Wave 2.4 | Agilent | https://www.agilent.com/en-us/products/cell-analysis-(seahorse)/seahorse-wave-software |
| CFX Manager 3.1 | Biorad | http://www.bio-rad.com/en-us/sku/1845000-cfx-manager-software?ID=1845000 |