KEY RESOURCES TABLE
| REAGENT or RESOURCE | SOURCE | IDENTIFIER |
|---|---|---|
| Antibodies | ||
|
| ||
| HA-Tag (C29F4) | Cell Signaling | 3724S; RRID:AB_1549585 |
| DPEAAE | Thermo Fisher | PA1-1748A; RRID:AB_2304324 |
| XCR1 (D2F8T) | Cell Signaling | 44665S; RRID:AB_2799269 |
| CLEC9A | Abcam | ab223188;RRID:AB_2884022 |
| CD8 (C8/144B) | eBioscience | 14-0085-80; RRID:AB_11151339 |
| c-myc (9E10)-chimeric | Novus Bio | NBP2-52636; RRID:AB_962987 |
| GAPDH– HRP | GenScript | A00192 |
| aPD-1 rat IgG2a (RPM1-14) | Bio X Cell InVivoPlus | BP0146; RRID:AB_10949053 |
| Isotype control (2A3) | Bio X Cell InVivoPlus | BE0089; RRID:AB_1107769 |
| NKp46/NCR1 | R&D | MAB1850; RRID:AB_2149153 |
| Anti-Asialo-GM1 | Wako Chemicals | NC1345696 |
| Flow cytometry antibodies are listed in Table S7 (sheet 1) | Table S7 (sheet 1) | Table S7 (sheet 1) |
| CyTOF antibodies are listed in Table S7 (sheet 3) | Table S7 (sheet 3) | Table S7 (sheet 3) |
|
| ||
| Bacterial and virus strains | ||
|
| ||
| pcDNA3-OVA | Addgene | 64599 |
| pHIV-Luc-ZsGreen | Addgene | 39196 |
| 5-alpha competent cells | New England Biolabs | C2987U |
| psPAX2 | Addgene | 12260 |
| pCMV-VSVg | Addgene | 8454 |
| shRNA clone against mouse Vcan | GeneCopoeia | MSH080253-LVRU6H |
| shRNA scrambled control clone for psi-LVRU6H | GeneCopoeia | CSHCTR001-LVRU6H |
|
| ||
| Biological samples | ||
|
| ||
| Human Lung Cancer TMA | US Biomax | BC041115e |
| Mouse subcutaneous tumor | This manuscript | N/A |
| Mouse lung | This manuscript | N/A |
| Mouse spine | This manuscript | N/A |
| Mouse long bones | This manuscript | N/A |
|
| ||
| Chemicals, peptides, and recombinant proteins | ||
|
| ||
| Ghost Dye 780 | Tonbo Biosciences | 13-0865 |
| DAPI | Sigma-Aldrich | D9542 |
| Collagenase Ia | Sigma-Aldrich | C2674 |
| Hyaluronidase V | Sigma-Aldrich | H6254 |
| Recombinant murine IL-6 | Peprotech | 216-16 |
| Recombinant murine GM-CSF | Peprotech | 315-03 |
| Recombinant human FLT3L | Peprotech | 300-19 |
| Chondroitinase ABC from Proteus vulgaris | Sigma-Aldrich | C2905 |
| Methanol | Fisher Scientific | A452-4 |
| TBS 10x | Corning | 46-012-CM |
| Tween20 | Sigma-Aldrich | P1379 |
| Clear-Rite 3 | Epredia | 6901 |
| 10x Tris/Glycine Buffer | Biorad | 1610734 |
| 10x Tris/Glycine/SDS Buffer | Biorad | 1610732 |
| Citrate-based Antigen Unmasking Solution | Vector Laboratories | H-3300 |
| Isoflurane USP | Midwest Vet Supply | NDC 13985-528-60 |
| 2-mercaptoethanol | Gibco | 21985-023 |
| DTT | Thermo Fisher | R0861 |
| 16% Formaldehyde Solution(w/v) | Thermo Scientific | TD264520 |
| Polybrene | Sigma-Aldrich | TR1003 |
| Hygromycin B | InvivoGen | ant-hg-1 |
| Blasticidin | InvivoGen | ant-bl-1 |
| LPS, E.Coli 0111:B4 | Sigma-Aldrich | LPS25 |
| DMXAA | InvivoGen | tlrl-dmx |
| Cell-ID Cisplatin | Fluidigm | 201064 |
| Cell-ID Intercalator-Ir | Fluidigm | 201192A |
| EQ beads | Fluidigm | 201078 |
| Staurosporine | Selleckchem | S1421 |
| OVA peptide 257-264 (SIINFEKL) | GenScript | RP10611 |
| Glycine 1M Solution | Sigma-Aldrich | 67419-1ML-F |
| UltraPure 0.5M EDTA | Invitrogen | 15575-038 |
| ECL Western Blotting Detection Reagents | Amersham | RPN2106 |
| Glutaraldehyde EM Grade 8% | Electron Microscopy Sciences | 2912.19.5000 |
| Cell Activation Cocktail with Brefeldin A | Biolegend | 423303 |
| Blotting grade blocker | Biorad | 1706404 |
| Bovine Serum Albumin (BSA) | Fisher bioreagents | BP 9703-100 |
| Isopropyl Alcohol | Sigma-Aldrich | PX1835-2 |
| DMSO | Sigma-Aldrich | D2650 |
| Phusion High Fidelity Polymerase | New England Biolabs | M0530S |
|
| ||
| Critical commercial assays | ||
|
| ||
| Wizard SV Genomic DNA Purification System | Promega | A2360 |
| GoTaq Master Mix | Promega | M7123 |
| RNAeasy Mini Kit | Qiagen | 74104 |
| Tissue Grinder System | Fisher Brand | 02-542-09 |
| NucleoBond Xtra Maxi kit | Macherey-Nagel | 740414.50 |
| Maxpar Antibody Labelling Kit | Fluidigm | 201160B |
| APC Annexin V Apoptosis Detection Kit with 7AAD | Biolegend | 640930 |
| Intracellular Fixation and Permeabilization Buffer | eBioscience | 88-8824-00 |
| AMPure XP | Beckman Coulter | A63880 |
| NEBNext® Ultra™ RNA Library Prep Kit for Illumina® | New England Biolabs | E7530L |
| NEBNext® RNA First Strand Synthesis Module | New England Biolabs | E7771 |
| Mouse Tumor Dissociation Kit | Miltenyi Biotec | 130-096-730 |
| Precision Count Beads | Biolegend | 424902 |
| RT2 Profiler PCR Array | Qiagen | PAMM-021Z |
| RT2 SYBR® Green qPCR Mastermix | Qiagen | 330529 |
| iSCript Reverse Transcription Supermix | Biorad | 1708840 |
| RT2 SYBR® Green qPCR Mastermix | Biorad | 330529 |
| SsoAdvanced Universal SYBR Green Supermix | Biorad | 1725272 |
| Quantikine mouse CXCL9 | R&D | MCX900 |
| Quantikine mouse IL27p28 | R&D | M2728 |
| Quantikine mouse IFNγ | R&D | P233156 |
| ToxinSensor Gel Clot Endotoxin Assay Kit | Genscript | L00351 |
| USER® Enzyme | New England Biolabs | M5508 |
| TruSeq PE Cluster Kit v3-cBot-HS | Illumina | PE-401-3001 |
|
| ||
| Deposited data | ||
|
| ||
| Raw and analyzed data | This manuscript | GEO: GSE199938 |
|
| ||
| Experimental models: Cell lines | ||
|
| ||
| Lewis Lung Carcinoma (LLC) | ATCC | CRL-1642 |
| B16F10 Melanoma | ATCC | CRL-6322 |
| 4T1 Mammary carcinoma | ATCC | CRL-2359 |
| MutuDC1940 | Applied Biological Materials | T0528 |
| VQ4935 | Wen et al., 2021 | N/A |
| HEK293T myc-versikine cells | Suneel S. Apte gift | N/A |
| HEK293T cells | ATCC | CRL-3216 |
|
| ||
| Experimental models: Organisms/strains | ||
|
| ||
| Mouse: C57BL/6J | The Jackson Laboratory | 000664 |
| Mouse: BALB/cJ | The Jackson Laboratory | 000651 |
| Mouse: B6.129 (Cg)-Cd44tm1Hbg/J | The Jackson Laboratory | 005085 |
| Mouse: B6.129S(C)-Batf3tm1Kmm/J | The Jackson Laboratory | 013755 |
| Mouse: B6.129-Tlr2tm1Kir/J | The Jackson Laboratory | 004650 |
| C57BL/6J-Tg(TcraTcrb), (OT-I) | The Jackson Laboratory | 003831 |
| VQ mice | Wen et al., 2021 | N/A |
|
| ||
| Oligonucleotides | ||
|
| ||
| Oligonucleotide qPCR primers are listed in Table S7 (sheet 2) | Table S7 (sheet 2) | Table S7 (sheet 2) |
| Oligonucleotide genotyping primers are listed in Table S7 (sheet 4) | Table S7 (sheet 4) | Table S7 (sheet 4) |
|
| ||
| Recombinant DNA | ||
|
| ||
| pLenti6-UbC-VKine-HA | Hope et al., 2016 | N/A |
| pLenti6-UbC-VKine-Myc | McCulloch et al., 2009 | N/A |
|
| ||
| Software and algorithms | ||
|
| ||
| FlowJo 10.7.1 | Tree Star | N/A |
| Graph Pad Prism Version:9.3.1 (350) | Dotmatics | N/A |
| Biorender | Biorender | N/A |
| SnapGene 6.0 | Dotmatics | N/A |
| OmniGraffle 7 | Omni Group | N/A |
| RT2 Profiler Data analysis | Qiagen | N/A |
|
| ||
| Other | ||
|
| ||
| Iscove’s DMEM medium | Corning | 15-016-CV |
| DMEM medium | Corning | 10-013-CV |
| RPMI 1640 medium | Corning | 10-040-CV |
| PBS | Corning | 21-040-CV |
| HEPES buffer 1M | Gibco | 15630-080 |
| Fetal Calf Serum One Shot | Gibco | A38400-01 |
| Antibiotic-antimycotic | Corning | 30-004-CI |
| Sodium Bicarbonate 7.5% (w/v) | Corning | 25-035-CI |
| MEM Non-essential amino acids | Gibco | 11140-050 |
| Glutamax | Gibco | 35050-061 |
| 0.25% Trypsin | Corning | 25-053-CI |
| BD Insulin Syringe | BD Insulin Syringe | 329461 |
| 5mL Polystyrene Round-Bottom Tube | Falcon | 352235 |
| Dual Chamber Cell Counting Slides | Biorad | 145-0011 |
| gentleMACS C Tubes | Miltenyi Biotec | 130-093-237 |
| MaxPar Cell Staining Buffer | Fluidigm | 201068 |
| MaxPar Fix and Perm Buffer | Fluidigm | 201067 |
| MaxPar Cell Acquisition Solution | Fluidigm | 201244 |
| Trypan Blue Stain (0.4%) | Gibco | 15250-061 |
| Red Blood Cell Lysing Buffer Hybri-Max | Sigma-Aldrich | R7757 |
| Mini-PROTEAN TGX Stain-Free Gels | Biorad | 4568096 |
| 2x Laemmli Sample Buffer | Biorad | 1610737 |
| Immobilon®-P PVDF Membrane | Millipore Sigma | IPVH15150 |
| PES 0.45 μm Filter | Cell Treat | 229748 |
| Sartorius Vivaspin 20 | Sartorius | VS2001 |
| Precision Plus Protein Dual Color Standards | Biorad | 161-0374 |
| Brilliant Stain Buffer | BD Biosciences | 563794 |
| UltraComp eBeads | Invitrogen | 01-2222-41 |