KEY RESOURCES TABLE.
| REAGENT or RESOURCE | SOURCE | DENTIFIER |
|---|---|---|
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| Antibodies | ||
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| In vivo anti-CD8, clone 2.43 | BioXcell | Cat# BE0061; RRID: AB_1125541 |
| In vivo anti-CD4, clone GK1.5 | BioXcell | Cat# BE0003–1; RRID: AB_1107636 |
| In vivo anti-IFNg antibody, clone XMG1.2 | BioXcell | Cat# BE0055; RRID: AB_1107694 |
| In vivo anti-CD25 antibody, clone PC-61.5.3 | BioXcell | Cat# BE0012; RRID: AB_1107619 |
| TruStain Fc (anti-mouse CD16/32) Antibody | BioLegend | Cat# 101320; RRID: AB_1574975 |
| APC/Fire 750 anti-CD3ε, clone 145–2C11 | BioLegend | Cat# 100362; RRID: AB_2629687 |
| BUV737 anti-CD4, clone RM4–5 | BD Biosciences | Cat# 612843; RRID: AB_2870165 |
| BV605 anti-CD8a, clone 53.6.7 | BioLegend | Cat# 100743; RRID: AB_2561352 |
| PE/Cyanine7 anti-CD11b, clone M1/70 | BioLegend | Cat# 101216; RRID: AB_312799 |
| BV650 anti-CD11c, clone N418 | BioLegend | Cat# 117339; RRID: AB_2562414 |
| Alexa Fluor 700 anti-CD19, clone 6D5 | BioLegend | Cat# 115528; RRID: AB_493735 |
| Alexa Fluor 488 anti-mouse CD45 Antibody, clone 30-F11 |
BioLegend | Cat# 103122; RRID: AB_493531 |
| BUV563 anti-CD103, clone M290 | BD Biosciences | Cat# 741261; RRID: AB_2870808 |
| BV711 anti-Gr-1, clone RB6–8C5 | BioLegend | Cat# 108443; RRID: AB_2562549 |
| BV786 Rat anti-mouse I-Ab/I-E, clone M5/114.15.2 | BD Biosciences | Cat# 742894; RRID: AB_2734759 |
| APC anti-mouse H-2Kb/H-2Db Antibody, clone 28–8-6 |
BioLegend | Cat# 114614; RRID: AB_2750194 |
| BV421 anti-FoxP3, clone MF-14 | BioLegend | Cat# 126419; RRID: AB_2565933 |
| PerCP/Cy5.5 anti-NK1.1, clone PK136 | BioLegend | Cat# 108728; RRID: AB_2132705 |
| PE anti-IFN-γ, clone XMG1.2 | Thermo Fisher | Cat# 12–7311-82; RRID: AB_466193 |
| PerCP-eFluor 710 anti-TNF-α, clone MP6-XT22 | Thermo Fisher | Cat# 46–7321-82; RRID: AB_1834445 |
| PE anti-CD274 (PD-L1) Antibody, clone 10F-9G2 | biolegend | Cat# 124308; RRID: AB_2073556 |
| PE anti-mouse CD366 (Tim3) antibody, clone RMT3–23 | BioLegend | Cat# 119704; RRID: AB_345378 |
| BV711 anti-mouse CD279 (PD-1) antibody, clone RMP1–30 | BioLegend | Cat# 135231; RRID: AB_2566158 |
| PE/Dazzle594 F4/80, clone BM8 | BioLegend | Cat# 123146; RRID: AB_2564133 |
| Monoclonal Anti-β-Actin antibody produced in mouse | Sigma-Aldrich, | Cat# A2228; RRID: AB_476697 |
| Stat1 (D1K9Y) Rabbit mAb | Cell Signaling | Cat# 14994; RRID: AB_2737027 |
| Stat3 (D1B2J) Rabbit mAb | Cell Signaling | Cat# 30835; RRID: AB_2798995 |
| Phospho-Stat1 (Tyr701) Rabbit, clone 58D6 | Cell Signaling | Cat# 9167; RRID: AB_561284 |
| Phospho-Stat3 (Tyr705) Antibody, clone D3A7 | Cell Signaling | Cat# 9145; RRID: AB_2491009 |
| anti-rabbit IgG HRP-linked antibody | Cell Signaling | Cat# 7074; RRID: AB_2099233 |
| anti-mouse IgG HRP-linked antibody | Cell Signaling | Cat# 7076;RRID: AB_330924 |
| Fatty Acid Synthase Antibody | Cell Signaling | Cat# 3189; RRID: AB_2100798 |
| Cleaved Caspase-3 (Asp175)Rabbit mAb, clone 5A1E | Cell Signaling | Cat# 9664; RRID: AB_2070042 |
| Recombinant Anti-p21 antibody, clone EPR18021 | Abcam | Cat# ab188224; RRID: AB_2734729 |
| Anti-CDKN2A/p16INK4a antibody | Abcam | Cat# ab211542; RRID: AB_2891084 |
| Recombinant Anti-Ki67 antibody, clone SP6 | Abcam | Cat# ab16667; RRID: AB_302459 |
| Recombinant Anti-c-Myc antibody, clone Y69 | Abcam | Cat# ab32072; RRID: AB_731658 |
| SLC23A2 (extracellular) Polyclonal Antibody | Invitrogen | Cat# PA5–111911; RRID: AB_2857319 |
| HIF-1 alpha (D1S7W) rabbit mAb | Cell Signaling | Cat# 36169; RRID: 2799095 |
| CD274 (PD-L1) Monoclonal Antibody, clone MIH5 | Invitrogen | Cat# 14–5982-82; RRID: AB_467781 |
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| Chemicals, peptides, and recombinant proteins | ||
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| Rotenone | Sigma-Aldrich | Cat#SI-R8875–1G |
| Antimycin A | Sigma-Aldrich | Cat#FL-75351–5MG |
| FCCP | Sigma-Aldrich | Cat#SI-C2920–10MG |
| Oligomycin A | Sigma-Aldrich | Cat#FL-75351–5MG |
| Collagenase | Sigma-Aldrich | Cat#C5138 |
| 4-Hydroxytamoxifen | Sigma-Aldrich | Cat# H7904–25MG |
| DMEM | GIBCO | Cat# 11965118 |
| RPMI 1640 Medium | GIBCO | Cat# 21875034 |
| 2-Mercaptoethanol | GIBCO | Cat# 31350010 |
| Murine IFN-g | Peprotech | Cat# 315–05 |
| ACK Buffer | Invitrogen | Cat# A10492–01 |
| Percoll | GE Healthca | Cat# 17–0891-01 |
| Dispase II | Roche (Sigma) | Cat#4942078001 |
| DnaseI | Sigma | Cat# D5025–150KU |
| True-Nuclear Transcription Factor Buffer Set | Biolegend | Cat#424401 |
| Permeabilization buffer 10X | eBioscience | Cat# 00–8333-56 |
| Stattic | Cayman | Cat# 14590 |
| Dynabeads Mouse T-Activator CD3/CD28 for T-Cell Expansion and Activation |
Invitrogen | Cat# 11452D |
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| Critical commercial assays | ||
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| Mouse XL Cytokine Array | R&D, | Cat#ARY028 |
| Senescence β-Galactosidase Staining Kit | Cell Signaling | Cat# 9860S |
| ApopTag Red In Situ Apoptosis Detection kit | Merck | Cat# S7165 |
| Hypoxyprobe Plus Kit | Hypoxyprobe | Cat# HP6–200Kit |
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| Deposited data | ||
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| RNA-seq data of cell lines | This paper | GEO: GSE217480 |
| ATAC-seq data of cell lines and primary tumor cells | This paper | GEO: GSE190993 and GSE217475 |
| RRBS data of cell lines | This paper | GEO: GSE21748 |
| Exome-seq data of cell lines | This paper | GEO: GSE217479 |
| TCR-seq of primary tumor cells | This paper | GEO: GSE217861 |
| RNA-seq data of primary tumor cells | This paper | GEO: GSE217862 |
| CRISPR screening | This paper | GEO: GSE217859 |
| Data and code availability | This paper | Data S1 |
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| Experimental models: Cell lines | ||
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| mouse: Yumm1.7 | ATCC | CRL-3362 |
| mouse: B16 | ATCC | CRL-6322 |
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| Experimental models: Organisms/strains | ||
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| Mouse: C57BL/6J | Jackson Laboratory | #:000664; RRID: IMSR_JAX:000664 |
| Mouse: Rag1−/− (B6.129S7-Rag1tm1Mom/J) | Jackson Laboratory | #:002216; RRID: IMSR_JAX:002216 |
| Mouse: mTmG (B6.129(Cg)-Gt(ROSA)26Sortm4(ACTB-tdTomato-EGFP)Luo/J) |
Jackson Laboratory | #:007676; RRID: IMSR_JAX:007676 |
| Mouse: IFNγ−/− (B6.129S7-Ifngtm1Ts/J) | Jackson Laboratory | #:002287; RRID: IMSR_JAX:002287 |
| Mouse: BRafCA; Tyr::CreER; Ptenlox4–5 (Braf/Pten) | Kindly provided by Dr. Marcus Bosenberg | N/A |
| Mouse: Kcre;Ecdf;P53f;RAG1 | Provided by Dr. Karin de Visser | N/A |
| Mouse: Neut;RAG1 | Provided by Dr. Karin de Visser | N/A |
| Mouse: Tyr::CreERT2/+; BRafCA/+; PtenVI; ROSA26RLSL-tdTomato/+ |
This paper | N/A |
| Mouse: Tyr::CreERT2/+; BRafCA/+; PtenVI; Rag1tm1Mom |
This paper | N/A |
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| Recombinant DNA | ||
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| pSCAR_sgRNA_puro-mKate-lox2272 | Addgene | Cat# 162076 |
| pSCAR_Cas9-hygro_GFP | Addgene | Cat# 162075 |
| pLX_EFS-Cre_ppt-del | Addgene | Cat# 162073 |
| psPAX2-D64V | Addgene | Cat# 63586 |
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| Software and algorithms | ||
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| Prism 9 | GraphPad | N/A |
| Flow Jo 10.1 | FlowJo | N/A |
| ImageJ | NIH | https://imagej.nih.gov |
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| Other | ||
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| LSRFortessa Flow Cytometer | BD Biosciences | N/A |
| LSRII Flow Cytometer | BD Biosciences | N/A |
| Seahorse XFe96 FluxPak | Agilent | Cat#102416–100 |
| Seahorse XFe96 Analyzer | Agilent | N/A |