KEY RESOURCES TABLE
| REAGENT or RESOURCE | SOURCE | IDENTIFIER |
|---|---|---|
| Antibodies | ||
| InVivoMab anti-mouse CD4 | BioXCell | Clone: GK1.5 Cat#: BE0003-1 |
| InVivoMab anti-mouse CD8α | BioXCell | Clone: 53-6.7 Cat#: BE0004-1 |
| InVivoMab anti-mouse F4/80 | BioXCell | Clone: CI:A3-1 Cat#: BE0206 |
| InVivoMab anti-mouse PD-1 | BioXCell | Clone: 29F.1A12 Cat#: BE0273 |
| InVivoMab anti-mouse ICOS | BioXCell |
Clone: 7E.17G9 Cat#: BE0059 |
| WB: Rabbit Ab Stat1 | Cell Signalling | Cat#: 9172 |
| WB: Rabbit mAb phosphor-Stat1 (Tyr701) | Cell Signalling | Cat#: 9167 |
| WB: Mouse mAb β-actin (8H10D10) | Cell Signalling | Cat#: 3700 |
| IHC/IF: anti-F4/80 antibody | Abcam | Clone: CI:A3-1 Cat#: ab6640 |
| IHC: anti-Arginase antibody | Abcam | Cat#: ab60176 |
| IHC: anti-CD3 antibody | Abcam | Clone: SP7 Cat#: ab16669 |
| IHC/IF: anti-RIP1 antibody | ProSci | Cat#: 5389 |
| IF: anti-CK19 antibody | DSHB at University of Iowa | Clone: Troma-III |
| IF: DAPI | Vector Labs | Cat#: H-1200 |
| Purified anti-mouse CD3ε | Biolegend | Clone: 145-2C11 |
| Purified anti-mouse CD28 | Biolegend | Clone: 37.51 |
| FC: anti-mouse CD44 | Biolegend | Clone: IM7 |
| FC: anti-mouse CD206 | Biolegend | Clone: C068C2 |
| FC: anti-mouse PD-1 | Biolegend | Clone: 29F.1A12 |
| FC: anti-mouse CD3 | Biolegend | Clone: 17A2 |
| FC: anti-mouse CD4 | Biolegend | Clone: RM4-5 |
| FC: anti-mouse CD8 | Biolegend | Clone: 53-6.7 |
| FC: anti-mouse CD45 | Biolegend | Clone: 30-F11 |
| FC: anti-mouse/human CD11b | Biolegend | Clone: M1/70 |
| FC: anti-mouse CD11c | Biolegend | Clone: N418 |
| FC: anti-mouse CD86 | Biolegend | Clone: GL-1 |
| FC: anti-mouse Gr1 | Biolegend | Clone: RB6-8C5 |
| FC: anti-mouse MHC II | Biolegend | Clone: M5/114.15.2 |
| FC: anti-mouse IL-10 | Biolegend | Clone: JES5-16E3 |
| FC: anti-mouse IFNγ | Biolegend | Clone: XMG1.2 |
| FC: anti-mouse TNFα | Biolegend | Clone: MP6-XT22 |
| FC: anti-mouse F4/80 | Biolegend | Clone: BM8 |
| FC: anti-mouse ICOS | Biolegend | Clone: 15F9 |
| FC: anti-mouse CD62L | Biolegend | Clone: MEL-14 |
| FC: anti-mouse CD25 | Biolegend | Clone: 3C7 |
| FC: anti-mouse CD40 | Biolegend | Clone: 3/23 |
| FC: anti-mouse CD69 | Biolegend | Clone: H1.2F3 |
| FC: anti-mouse IL-17A | Biolegend | Clone: TC11-18H10.1 |
| FC: anti-mouse TGFβ | Biolegend | Clone: TW7-16B4 |
| FC: anti-mouse LFA-1 | Biolegend | Clone: H155-78 |
| FC: anti-mouse LAG-3 | Biolegend | Clone: C9B7W |
| FC: anti-STAT1 Phospho (Ser727) | Biolegend | Clone: A15158B |
| FC: anti-STAT3 Phospho (Tyr705) | Biolegend | Clone: 13A3-1 |
| FC: anti-STAT6 Phospho (Tyr641) | Biolegend | Clone: A15137E |
| FC: anti-STAT5 Phospho (pY694) | BD Biosciences | Clone: 47/Stat5(pY694) |
| FC: anti-mouse T-bet | eBioscience | Clone: eBio4B10 |
| FC: anti-mouse RORγt | eBioscience | Clone: AFKJS-9 |
| FC: anti-mouse Granzyme B | eBioscience | Clone: NGZB |
| FC: anti-mouse Perforin | eBioscience | Clone: eBioOMAK-D |
| FC: anti-mouse FoxP3 | eBioscience | Clone: FJK-16s |
| FC: anti-human CD45 | Biolegend | Clone: HI30 |
| FC: anti-human CD3 | Biolegend | Clone: UCHT1 |
| FC: anti-human CD4 | Biolegend | Clone: A161A1 |
| FC: anti-human CD25 | Biolegend | Clone: BC96 |
| FC: anti-human CD69 | Biolegend | Clone: FN50 |
| FC: anti-human CD11b | Biolegend | Clone: ICRF44 |
| FC: anti-human TNFα | Biolegend | Clone: Mab11 |
| FC: anti-human IFNγ | Biolegend | Clone: 4S.B3 |
| FC: anti-human IL-10 | Biolegend | Clone: JES3-19F1 |
| FC: anti-human HLA-DR | Biolegend | Clone: L243 |
| Bacterial and Virus Strains | ||
| Biological Samples | ||
| Patient-derived organotypic tumor spheroids | NYU Langone Medical Center | N/A |
| Chemicals, Peptides, and Recombinant Proteins | ||
| Recombinant Mouse TNF-alpha | Cell Sciences | Cat#: CRT192C |
| Recombinant Human M-CSF Protein | R&D | Cat#: 216-MC |
| zVAD-FMK | Bachem | Cat#: N-1510 |
| Nec-1s | BioVision | Cat#: 2263 |
| GSK’963 | GlaxoSmithKline | N/A |
| RIP1i (GSK’547) | GlaxoSmithKline | N/A |
| Fludarabine | Selleckchem | S1491; CAS: 21679-14-1 |
| LJI308 | Sigma-Aldrich | SML1788; CAS: 1627709-94-7 |
| GW9662 | Sigma-Aldrich | M6191; CAS: 22978-25-2 |
| Albumin-fluorescein isothiocyanate conjugate | Sigma-Aldrich | A9771; MDL: MFCD00282182 |
| Rodent Chow (Purina chow 5001) formulated with inhibitor | Research Diets | Cat#: C16102601i |
| QVD-Oph | Millipore Sigma | 551476 |
| Recombinant human TNFα | R&D systems | 210-TA-100 |
| FLAG-RIPK1 1-294 (C34A C127A C233A C240A) | Genscript | N/A |
| fluorescently-labeled ligand (14-(2-{[3-({2-{[4-(cyanomethyl)phenyl]amino}-6-[(5-cyclopropyl-1H-pyrazol-3-yl)amino]-4-pyrimidinyl}amino) propyl]amino}-2-oxoethyl)-16,16,18,18-tetramethyl-6,7,7a,8a,9,10,16,18-octahydrobenzo [2’,3’]indolizino[8’,7’:5′,6′]pyrano [3′,2′:3,4]pyrido[1,2-a]indol-5-ium-2-sulfonate) | GSK | N/A |
| Critical Commercial Assays | ||
| MILLIPLEX Human Cytokine 41-Plex kit | MilliporeSigma | Cat#: HCYTMAG-60K-PX41 |
| Reaction Biology Kinase Assay | Reaction Biology Corp | Wild type kinase panel |
| Cell Titre-Glo Luminescent Cell Viability Assay | Promega | G7573 |
| Deposited Data | ||
| X-ray crystalllography data, maps and structure coordinates: PDB | This paper | 6HHO https://www.rcsb.org/ |
| Raw and analyzed RNAseq data | This paper | GEO: GSE111626 |
| Experimental Models: Cell Lines | ||
| Mouse: KPC Pancreatic Cancer Cell line | Laboratory of Dave Tuveson | Cell line: FC1242 |
| L929 (Mouse L-cells NCTC 929) | ATCC | ATCC CCL-1 |
| Experimental Models: Organisms/Strains | ||
| Mouse: C57BL/6 (H-2Kb) | The Jackson Laboratory | JAX: 000664 |
| Mouse: C57BL/6-Tg(TcraTcrb)1100Mjb/J (OT-I) | The Jackson Laboratory | JAX: 003831 |
| Mouse: B6.Cg-Tg(TcraTcrb)425Cbn/J (OT-II) | The Jackson Laboratory | JAX: 004194 |
| Mouse: B6.129S(Cg)-Stat1tm1Dlv/J | The Jackson Laboratory | JAX: 012606 |
| Mouse: B6.129S7-Rag1tm1Mom/J | The Jackson Laboratory | JAX: 002216 |
| Mouse: B6.Cg-Foxn1nu/J (B6 nude) | The Jackson Laboratory | JAX: 000819 |
| Mouse: Ripk3−/− | Genentech | N/A |
| Mouse: RIP1 KD/KI | GlaxoSmithKline | N/A |
| Mouse: B6. LSL-KrasG12D and p48Cre (KC ) | Laboratory of Dafna Bar-Sagi | N/A |
| Oligonucleotides | ||
| Primers for qPCR, see Table S5 | This paper | N/A |
| Recombinant DNA | ||
| RIP1 shRNA Plasmid: SHCLNG-NM_009068 | Sigma-Aldrich | TRCN0000022466 |
| RIP1 shRNA Plasmid: SHCLNG-NM_009068 | Sigma-Aldrich | TRCN0000278133 |
| Software and Algorithms | ||
| GraphPad Prism 7 | GraphPad Software | |
| Analyst multimode reader | Molecular Devices | 0200 |
| iMOSFLM | CCP4 | N/A |
| Aimless | CCP4 | 0.512 |
| ctruncate | CCP4 | 1.16.9 |
| unique | CCP4 | 6.5.013 |
| freerflag | CCP4 | 1.2 |
| pointless | CCP4 | 1.9.16 |
| Other | ||