REAGENT or RESOURCE | SOURCE | IDENTIFIER |
---|---|---|
Antibodies | ||
Anti-PD-L1 (mouse IgG2A, clone 6E11) | Genentech | N/A |
Anti-IL6R (mouse IgG2A, clone MR16-1) | Genentech | N/A |
Mouse IgG2A isotype control (anti-gp120) | Genentech | N/A |
Anti-mouse-DEC205-ovalbumin | Genentech | N/A |
Anti-mouse-CD40 (clone FGK4.5) | Genentech | N/A |
Rabbit anti-phospho-STAT3 (Y705; clone D3A7) | Cell Signaling Technology | Cat#9145; RRID:AB_2491009 |
Rabbit anti-STAT3 (clone D1B2J) | Cell Signaling Technology | Cat#30835; RRID:AB_2798995 |
Mouse anti-β-actin-HRP (clone AC-15) | Sigma-Aldrich | Cat#A3854; RRID:AB_262011 |
Rabbit anti-BATF (clone D7C5) | Cell Signaling Technology | Cat#8638; RRID:AB_11141425 |
Hamster anti-mouse CD3e (clone 145-2C11) | BD Biosciences | Cat#550275; RRID:AB_393572 |
Hamster anti-mouse CD28 (clone 37.51) | BD Biosciences | Cat#553295; RRID:AB_394764 |
Rat anti-mouse CD16/32 (clone 2.4G2) | BD Biosciences | Cat#553142; RRID:AB_394657 |
Anti-mouse CD45-BV510 (clone 30-F11) | BD Biosciences | Cat#563891; RRID:AB_2734134 |
Anti-mouse Thy1.2-efluor450 (clone 53-2.1) | eBioscience | Cat#48-0902-82; RRID:AB_1272200 |
Anti-mouse Thy1.2-alexafluor700 (clone 53-2.1) | Biolegend | Cat#140324; RRID:AB_2566740 |
Anti-mouse Thy1.1-alexafluor488 (clone OX-7) | Biolegend | Cat#202506; RRID:AB_492882 |
Anti-mouse CD3ε-PE/Cy5.5 (clone 145-2C11) | Biolegend | Cat#100310; RRID:AB_312675 |
Anti-mouse CD4-BUV395 (clone GK1.5) | BD Biosciences | Cat#563790; RRID:AB_2738426 |
Anti-mouse CD4-BV785 (clone GK1.5) | Biolegend | Cat#100453; RRID:AB_2565843 |
Anti-mouse CD8a-BB515 (clone 53-6.7) | BD Biosciences | Cat#564422; RRID:AB_2738801 |
Anti-mouse CD8a-PE (clone 53-6.7) | Biolegend | Cat#100708; RRID:AB_312747 |
Anti-mouse CD8a-BUV737 (clone 53-6.7) | BD Biosciences | Cat#612759; RRID:AB_2870090 |
Anti-mouse CD11b-alexafluor700 (clone M1/70) | Biolegend | Cat#101222; RRID:AB_493705 |
Anti-mouse Gr1-PE-Cy5.5 (clone RB6-8C5) | eBioscience | Cat#35-5931-82; RRID:AB_469740 |
Anti-mouse CD11c-PE/Dazzle594 (clone N418) | Biolegend | Cat#117348; RRID:AB_2563655 |
Anti-mouse MHCII (I-A/I-E)-FITC (clone M5/114.15.2) | eBioscience | Cat#11-5321-85; RRID:AB_465233 |
Anti-mouse CD64-PE/Cy7 (clone X54-5/7.1) | Biolegend | Cat#139314; RRID:AB_2563904 |
Anti-mouse CD169-PE/Cy7 (clone 3D6.112) | Biolegend | Cat#142412; RRID:AB_2563911 |
Anti-mouse B220-BUV737 (clone RA3-6B2) | BD Biosciences | Cat#612838; RRID:AB_2870160 |
Anti-mouse IL6R-PE (clone D7715A7) | Biolegend | Cat#115806; RRID:AB_313677 |
Anti-mouse gp130-PE (clone 4H1B35) | Biolegend | Cat#149403; RRID:AB_2566644 |
Anti-mouse Foxp3-efluor450 (clone FJK-16s) | eBioscience | Cat#48-5773-82; RRID:AB_1518812 |
Anti-mouse/human GzmB-Pacific blue (clone GB11) | Biolegend | Cat#515408; RRID:AB_2562196 |
Anti-mouse TNF-PE (clone MP6-XT22) | Biolegend | Cat#506306; RRID:AB_315427 |
Anti-mouse IFNγ-PE/Dazzle594 (clone XMG1.2) | Biolegend | Cat#505846; RRID:AB_2563980 |
Anti-mouse IL-17A-BV785 (clone C11-18H10) | Biolegend | Cat#506928; RRID:AB_2629787 |
Anti-human CD8a-BUV395 (clone RPA-T8) | BD Biosciences | Cat#563796 |
Anti-human TNF-alexafluor488 (clone MAb11) | Biolegend | Cat#502917; RRID:AB_493122 |
Anti-human IFNγ-APC (clone B27) | Biolegend | Cat#506510; RRID:AB_315443 |
Anti-human CD3 (clone OKT3) | eBioscience | Cat#16-0037-85; RRID:AB_468855 |
Anti-human CD28 (clone CD28.2) | BD Biosciences | Cat#555725; RRID:AB_396068 |
Biological samples | ||
Pre-treatment PBMC from Atezolizumab trials (IMmotion150 and IMvigor210) | Genentech | N/A |
Healthy donor PBMC | Genentech | N/A |
Renal cell cancer tissue blocks from IMmotion150 | Genentech | N/A |
Chemicals, peptides, and recombinant proteins | ||
TrueCut Cas9 Protein v2 | Thermo Fisher Scientific | Cat#A36499 |
Recombinant mouse IL-6 | R&D Systems | Cat#406-ML |
Recombinant mouse hyper-IL-6 | R&D Systems | Cat# 9038-SR-025/CF |
Recombinant human IL-6 | R&D Systems | Cat#206-IL |
Recombinant human IL-2 | R&D Systems | Cat#202-IL |
SIINFEKL (OVA 257-264) peptide | AnaSpec | Cat#AS-60193-1 |
Fixable Viability Dye eFluor™ 780 | Invitrogen | Cat#65-0865-14 |
Cell Stimulation Cocktail plus protein transport inhibitors (500X) | Invitrogen | Cat#00-4975-03 |
Brefeldin A (1000X) | eBioscience | Cat#00-4506-51 |
Critical commercial assays | ||
Foxp3/Transcription Factor Staining Buffer Set | eBioscience | Cat#00-5523-00 |
EasySep Mouse CD8+ T cell Isolation Kit | StemCell Technologies | Cat#19853 |
EasySep Mouse Naive CD8+ T cell Isolation Kit | StemCell Technologies | Cat#19858 |
EasySep Human Naive CD8+ T cell Isolation Kit | StemCell Technologies | Cat#19258 |
RNEasy Mini Kit | Qiagen | Cat#74104 |
iScript cDNA synthesis kit | Biorad | Cat#1708891 |
Deposited data | ||
Raw and analyzed bulk RNAseq data (mouse) | This paper | GEO: GSE199048 |
Tumor expression data (IMmotion150) | EGA | EGA: EGAS00001004387 |
Human PBMC single-cell RNAseq count data | EGA | EGA: EGAS00001004451, EGAS00001004458 |
Experimental models: Cell lines | ||
EMT6 | Genentech | N/A |
MC38 | Genentech | N/A |
MC38-GFP | Genentech | N/A |
CT26 | Genentech | N/A |
EG7.OVA | Genentech | N/A |
Experimental models: Organisms/strains | ||
Mouse: C57BL/6J | The Jackson Laboratory | JAX: 000664 |
Mouse: BALB/c | Charles River | Model# 028 |
Mouse: B6; SJL-Il6ratm1.1Drew/J | The Jackson Laboratory | JAX: 012944 |
Mouse: Rosa26-Cre.ki (C57BL/6NTac-Gt(ROSA)26Sortm16(cre)Arte) | Taconic | Model# 12524 |
Mouse: IL6R−/− (B6; SJL-Il6ratm1.1Drew/J x C57BL/6NTac-Gt(ROSA)26Sortm16(cre)Arte | This paper | N/A |
Mouse OT-I: C57BL/6Tg(TcraTcrb)1100Mjb/J | The Jackson Laboratory | JAX: 003831 |
Mouse: IL6R−/− OT-1 (C57BL/6Tg(TcraTcrb)1100Mjb/J x C57BL/6Il6r−/−) | This paper | N/A |
Mouse: Cd4-Cre (B6.Cg-Tg(CD4-cre)1Cwi N9) | Taconic | Model# 4196 |
Mouse: Stat3flox (B6.129S1-Stat3tm1Xyfu/J) | The Jackson Laboratory | JAX: 016923 |
Mouse: CD4ΔSTAT3 (B6.Cg-Tg(CD4-cre)1Cwi N9 x B6.129S1-Stat3tm1Xyfu/J) | This paper | N/A |
Mouse: Cd8a-Cre (C57BL/6-Tg(Cd8a-cre)1Itan/J) | The Jackson Laboratory | JAX: 008766 |
Mouse: CD8ΔIL6R (C57BL/6-Tg(Cd8a-cre)1Itan/J x B6; SJL-Il6ratm1.1Drew/J) | This paper | N/A |
Oligonucleotides | ||
Rpl19 (Mm02601633_g1) | Life Technologies | Cat#4331182 |
Il6ra (Mm01211445_m1) | Life Technologies | Cat#4331182 |
Il6st (Mm00439665_m1) | Life Technologies | Cat#4331182 |
Batf (Mm00479410_m1) | Life Technologies | Cat#4331182 |
Guide targeting exon 1 of Batf 5′ GGGGGTACCTGTTTGCCAG-3′ | IDT | N/A |
Alt-R CRISPR-Cas9 tracrRNA | IDT | Cat#1072534 |
Alt-R CRISPR-Cas9 negative control crRNA | IDT | Cat#1072544 |
IL6 in situ hybridization probe | Advanced Cell Diagnostics | Cat#2-1082 |
Software and algorithms | ||
FlowJo | FlowJo | https://www.flowjo.com/ |
Prism 9 | GraphPad | https://www.graphpad.com/ |
R | The R Project for Statistical Computing | https://www.r-project.org/ |
Microsoft Excel for Mac | Microsoft | https://products.office.com/en-us/excel |