KEY RESOURCES TABLE
| REAGENT or RESOURCE | SOURCE | IDENTIFIER |
|---|---|---|
|
| ||
| Antibodies | ||
|
| ||
| αCD40 (Clone FGK45) | JPP Biologics | N/A |
| αPD1 (Clone RMP1–14) | JPP Biologics | N/A |
| αCTLA4 (Clone 4F10) | JPP Biologics | N/A |
| αCD4 (Clone GK1.5) | JPP Biologics | N/A |
| αCD8 (Clone YTS169) | JPP Biologics | N/A |
| αNK1.1 (Clone PK136) | JPP Biologics | N/A |
| αCSF1R (Clone AFS98) | BioxCell | Cat#BP0213 |
| αIL12 (Clone R2–9A5) | BioxCell | Cat#BE0233 |
| αCXCR3 (Clone CXCR3–173) | BioxCell | Cat#BE0249 |
| Fc block | eBioscience | Cat#14–9161-73 |
| APC/Cyanine7 rat anti-mouse CD45 (Clone 30-F11) | BioLegend | Cat#103116 |
| PerCP/Cyanine5.5 rat anti-mouse CD45 Antibody (Clone 30-F11) | BioLegend | Cat#103131 |
| APC mouse anti-mouse NK1.1 (Clone PK136) | eBioscience | Cat#17–5941-82 |
| PE rat anti-mouse TCR β chain (Clone H57–597) | BioLegend | Cat#109208 |
| CD8a (Clone 53–6.7) | BioLegend | Cat#100722 |
| FITC rat anti-mouse F4/80 (Clone BM8) | BioLegend | Cat#123108 |
| PE-Cy7 rat anti-mouse F4/80 (Clone BM8) | eBioScience | Cat#25–4801-82 |
| PE rat anti-mouse CD11b (Clone M1/70 | BD Biosciences | Cat#553311 |
| PerCP-Cyanine5.5 rat anti-mouse Ly6G (Clone 1A8) | BD Biosciences | Cat#560602 |
| eFluor 450 rat anti-mouse Ly6C (Clone HK1.4) | eBioscience | Cat#48–5932-82 |
| APC rat anti-mouse Ly6C (Clone HK1.4) | BioLegend | Cat#128016 |
| APC rat anti-mouse CD206 (Clone C068C2) | BioLegend | Cat#141708 |
| FITC rat anti-mouse CD11c (Clone 3.9) | eBioscience | Cat#11–0116-42 |
| eFluor 450 rat anti-mouse MHC II (Clone M5/114.15.2) | eBioscience | Cat#48–5321-82 |
| Brilliant Violet 785 rat anti-mouse MHC II (Clone M5/114.15.2) | BioLegend | Cat#107645 |
| PE hamster anti-mouse CD103 (Clone 2E7) | eBioscience | Cat#12–1031-82 |
| PE rat anti-mouse PDGFRα (Clone APA5) | BioLegend | Cat#135906 |
| Pe-Cy7 rat anti-mouse CD31 (Clone 390) | BioLegend | Cat#102418 |
| APC-Cy7 hamster anti-mouse PDPN (Clone 8.1.1) | BioLegend | Cat#127418 |
| FITC rat anti-mouse EpCAM (Clone G8.8) | BioLegend | Cat#118208 |
| Rabbit anti-mouse Fibronectin | Abcam | Cat#ab2143 |
| Rabbit anti-mouse PDPN | Abcam | Cat#ab11936 |
| Rabbit anti-mouse αSMA | Abcam | Cat#ab5694 |
| Rabbit anti-mouse PDGFRβ | Abcam | Cat#ab32570 |
| Rat anti-mouse CD8a (Clone 4SM15) | eBioscience | Cat#14–0808-82 |
| Rabbit anti-mouse Granzyme B (Clone D6E9W) | Cell Signaling Technology | Cat#44153 |
| Rabbit anti-mouse Perforin (Clone E3W4I) | Cell Signaling Technology | Cat#31647 |
| Rabbit anti-rat biotinylated IgG (Avidin Biotin Complex kit) | Vector Laboratories | Cat#BA-4001 |
| Goat anti-rabbit biotinylated IgG (Avidin Biotin Complex kit) | Vector Laboratories | Cat#BA-1000 |
|
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| Critical commercial assays | ||
|
| ||
| RN-easy Micro Plus kit | Qiagen | Cat#74034 |
| RN-easy Mini Plus kit | Qiagen | Cat#74134 |
| SuperScript™ IV First-Strand Synthesis System | Invitrogen | Cat#18091050 |
| TaqMan™ Fast Universal PCR Master Mix (2X), no AmpErase™ UNG | Applied Biosystems | Cat#4352042 |
|
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| Deposited data | ||
|
| ||
| Bulk RNA sequencing data of KPC liver metastases from WT and HckKO mice | This paper | GEO repository GSE185540 |
| Single cell RNA sequencing of human pancreatic cancer and mouse KPC tumors | Elyada et al., 2019 |
Elyada et al., 2019 PMID: 31197017 |
|
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| Experimental models: Cell lines | ||
|
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| Mouse KPC pancreatic cancer cells | A/Prof Marina Pajic, The Garvan Institute of Medical Research, Australia. |
Vennin et al., 2017 PMID: 28381539 |
|
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| Experimental models: Organisms/strains | ||
|
| ||
| Mouse: Wild-type C57BL/6 | In-house (Prof Matthias Ernst), The Olivia Newton-John Cancer Research Institute. | N/A |
| Mouse: HckKO | In-house (Prof Clifford Lowell), The Olivia Newton-John Cancer Research Institute. |
Lowell et al. (1994) PMID: 8125254 |
| Mouse: Rag1KO | In-house (Prof Matthias Ernst), The Olivia Newton-John Cancer Research Institute. | N/A |
| Mouse: HckKO;Rag1KO | In-house (Prof Matthias Ernst), The Olivia Newton-John Cancer Research Institute. | N/A |
| Mouse: ItgaxCreIrf8fl/fl | In-house (Dr Michael Chopin), The Walter and Eliza Hall Institute, Australia. |
Chopin et al. (2013) PMID: 24249112 |
|
| ||
| Oligonucleotides | ||
|
| ||
| See Table S12 for Taqman Probes | N/A | |
|
| ||
| Software and algorithms | ||
|
| ||
| Aperio ImageScope v11.2.0.780 | Leica Biosystems | https://www.leicabiosystems.com/en-au/digital-pathology/manage/aperio-imagescope/ |
| FIJI (ImageJ) |
Schneider et al. (2012) PMID: 22930834 |
https://imagej.nih.gov/ij/ |
| FlowJo (Version 10) | Flowjo | https://www.flowjo.com/ |
| GraphPad Prism Software (Version 8) | GraphPad | https://www.graphpad.com/scientific-software/prism/ |
| Subread aligner (Rsubread version 2.2.6) |
Liao et al., 2013, 2019 PMIDs: 23558742 & 30783653 |
https://bioconductor.org/packages/release/bioc/html/Rsubread.html |
| featureCounts (Rsubread version 2.2.6) |
Liao et al. 2014, 2019 PMIDs: 24227677 & 30783653 |
https://bioconductor.org/packages/release/bioc/html/Rsubread.html |
| limma (3.44.3) |
Law et al. (2014) PMID: 24485249 Ritchie et al. (2015) PMID: 25605792 |
https://bioconductor.org/packages/release/bioc/html/limma.html |
| Seurat (version 4) |
Hao et al. (2021) PMID: 34062119 |
https://cran.r-project.org/web/packages/Seurat/index.html |
| R version 4.0.3 | CRAN | https://cran.csiro.au/src/base/R-4/R-4.0.3.tar.gz |
| cellCounts (Rsubread version 2.2.6) |
Liao et al., 2013, 2019 PMIDs: 23558742 & 30783653 |
https://bioconductor.org/packages/release/bioc/html/Rsubread.html |
|
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| Other | ||
|
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| DMEM/F-12 media | Gibco | Cat#11320033 |
| HBSS media | Gibco | Cat#14170112 |
| Ketamine | Baxter | N/A |
| Xylazine | Ilium | N/A |
| Isoflurane | Baxter | N/A |
| Carprofen | Zoetis | N/A |
| Bupivacaine | AstraZeneca | N/A |
| Gemcitabine | Pfizer | N/A |
| Neomycin | ThermoFisher | Cat#21810031 |
| RK20449 | Reagency | Saito et al. (2013) PMID: 23596204 |
| Captisol | Captisol | Cat#RC-0C7-K01 |
| Collagenase / Dispase | Roche | Cat#11097113001 |
| DNase I | Roche | Cat#10104159001 |
| Sytox Blue | Invitrogen | Cat#S34857 |
| Normal goat serum | Gibco | Cat#PCN5000 |