KEY RESOURCES TABLE
| REAGENT or RESOURCE | SOURCE | IDENTIFIER |
|---|---|---|
| Antibodies | ||
| Anti-mouse CD45, PerCp-Cy5.5 conjugated, clone:30-F11 | BD PharMingen | Cat# 550994; RRID:AB_394003 |
| Anti-mouse CD3e, FITC conjugated, clone: 145–2C11 | BD PharMingen | Cat# 553061; RRID:AB_394594 |
| Anti-mouse CD8a, BV510 conjugated, clone: 53-6.7 | BD PharMingen | Cat# 563068; RRID:AB_2687548 |
| Anti-mouse CD4, APC conjugated, clone: RM4-5 | BD PharMingen | Cat# 553051; RRID:AB_398528 |
| Anti-mouse Foxp3, PE conjugated, clone: MF23 | BD PharMingen | Cat# 560408; RRID:AB_1645251 |
| Anti-mouse CD11b, PE conjugated, clone: M1/70 | BD PharMingen | Cat# 557397; RRID:AB_396680 |
| Anti-mouse F4/80, APC conjugated, clone: BM8 | BioLegend | Cat # 123116; RRID:AB_893481 |
| Anti-mouse Ly-6G, FITC conjugated, clone: 1A8 | BD PharMingen | Cat# 551460; RRID:AB_394207 |
| Anti-mouse CD16/CD32 (Fc block), clone: 2.4G2 | BD PharMingen | Cat # 553142; RRID: AB_394657 |
| Anti-mouse CD3e, PerCP-Cy5.5 conjugated, clone: 145–2C11 | BD PharMingen | Cat# 551163; RRID:AB_394082 |
| Anti-mouse CD8a, FITC conjugated, clone: 53-6.7 | BD PharMingen | Cat# 553031; RRID:AB_394569 |
| Anti-mouse CD4, FITC conjugated, clone: RM4-5 | BD PharMingen | Cat# 553047; RRID:AB_394583 |
| Anti-mouse CD11c, PE conjugated, clone: HL3 | BD PharMingen | Cat# 553802; RRID:AB_395061 |
| Anti-mouse CD103, BV421 conjugated, clone: M290 | BD PharMingen | Cat # 562771; RRID: AB_2737783 |
| Anti-mouse F4/80, FITC conjugated, clone: BM8 | BioLegend | Cat # 123108; RRID: AB_893502 |
| Anti-mouse B220, FITC conjugated, clone: RA3–6B2 | BioLegend | Cat # 103206; RRID: AB_312991 |
| Anti-mouse I-A/I-E (MHCII) Antibody, PE-Cy7 conjugated, clone: M5/114.15.2 | BioLegend | Cat # 107628; RRID: AB_2069377 |
| Anti-mouse CD8a, APC conjugated, clone: 53-6.7 | BD PharMingen | Cat# 553035; RRID: AB_398527 |
| Rabbit anti-mouse CD8a, clone: D4W2Z | Cell Signaling Technologies | Cat# 98941; RRID:AB_2756376 |
| Anti-mouse CXCL5 | LsBio | Cat # LS-C104413; RRID AB_10624103 |
| Rat anti-mouse Ly6G | Abcam | Cat# ab25377; RRID:AB_470492 |
| Rat anti-moue IDO1, clone: mIDO-48 | Santa Cruz Biotechnology | Cat# sc-53978; RRID:AB_831071 |
| Rabbit anti-TDO2 | Proteintech | Cat# 15880–1-AP; RRID:AB_2827610 |
| Rabbit anti-mouse β-Catenin, clone: D10A8 | Cell Signaling Technologies | Cat# 8480; RRID:AB_11127855 |
| Anti-mouse CD45, PerCp-Cy5.5 conjugated, clone:30-F11 | BD PharMingen | Cat# 550994; RRID:AB_394003 |
| Anti-mouse CD3e, FITC conjugated, clone: 145–2C11 | BD PharMingen | Cat# 553061; RRID:AB_394594 |
| Anti-mouse CD8a, BV510 conjugated, clone: 53-6.7 | BD PharMingen | Cat# 563068; RRID:AB_2687548 |
| OMP-18R5 | OncoMed | N/A |
| OMP-54F28 | OncoMed | N/A |
| Anti-PD-1, clone: RMP-14 | BioXCell | Cat# BE0146; RRID:AB_10949053 |
| Anti-CD8 | Duke Cell Culture Facility | N/A |
| Rat IgG2a isotype control, clone: 2A3 | BioXCell | Cat# BE0089; RRID: AB_1107769 |
| Bacterial and virus strains | ||
| Ad5CMVCre | Univ. of Iowa | Cat # Ad5CMVCre |
| Biological Samples | ||
| Human melanoma tissue specimens | Duke University Hospital | N/A |
| Pre and Post ETC-159 treated human tumor specimens | A*STAR | N/A |
| Chemicals, peptides, and recombinant proteins | ||
| Collagenase IV | Sigma-Aldrich | Cat # C-5138 |
| Hyaluronidase | Sigma-Aldrich | Cat # H-6254 |
| DNaseI | Sigma-Aldrich | Cat # D-5025 |
| RPMI | Sigma-Aldrich | Cat # R8758 |
| RBC Lysis Buffer | Sigma-Aldrich | Cat # R7757 |
| Live/Dead Fixable Violet Dead Cell Stain Kit | ThermoFisher | Cat # L34955 |
| Live/Dead Fixable Aqua Dead Cell Stain Kit | ThermoFisher | Cat # L34966 |
| Recombinant Wnt5a | R&D systems | Cat # 645-WN-010 |
| Aminolevulenic acid | Sigma | Cat # A3785 |
| Hematoxylin | VWR | Cat # 95057–844 |
| Eosin | VWR | Cat # 95057–848 |
| Vina Green Chromogen Kit | BioCare Medical | Cat # BRR 807 AH |
| Warp Red Chromogen Kit | BioCare Medical | Cat # 901-WR806–081017 |
| NP40 lysis buffer | Sigma-Aldrich | Cat #492016 |
| RIPA Lysis and Extraction Buffer | ThermoFisher | Cat # 89901 |
| Protease Inhibitor Cocktail | Roche | Cat #4693159001 |
| PhosSTOP | Roche | Cat # 4906845001 |
| TRP2180–188 peptide | ANASPEC | Cat # AS-61058 |
| MUT151–58 peptide | BACHEM | Cat # 4026648 |
| OVA257–264 peptide | InvivoGen | Cat # vac-sin |
| C-59 | Selleckchem | Cat # S7037 |
| ETC-159 | A*STAR | N/A |
| Collagenase IV | Sigma-Aldrich | Cat # C-5138 |
| Recombinant Mouse IL-4 | BioAbChem | Cat # 42-IL4 |
| Recombinant Mouse GM-CSF | R&D System | Cat # 415-ML-010 |
| 4-hydroxytamoxifen | Sigma-Aldrich | Cat # H6278–10MG |
| Polyethyelene glycol(PEG)-8000 | Sigma-Aldrich | Cat # 89510–250G-F |
| Critical commercial assays | ||
| Mouse Kynurenine ELISA kit | MyBioSource | Cat # MBS043489 |
| Mouse IFNγ ELISPOT Plus | Mabtech | Cat # 3321-4APT-2 |
| Mouse FoxP3 Buffer Set | BD Biosciences | Cat # 560409 |
| Miltenyi CD11c magnetic selection columns | Miltenyi Biotec | Cat # 130-108-338 |
| Naive CD4+ T cells selection kit | Stem Cell Technologies | Cat # 19765 |
| RNAeasy Plus Micro kit | QIAGEN | Cat # 74034 |
| SuperScript IV FirstStrand Synthesis System | Invitrogen | Cat # 11756050 |
| PowerUp Master Mix | Applied Biosciences | Cat # A25742 |
| Deposited data | ||
| RNA-seq data derived from the murine BRAFV600EPTEN−/− melanoma anti-PD-1 resistance study | Hanks Lab; Theivanthiran et al., 2020; Zhao et al., 2018a | SAMN09878780 |
| RNA-seq dataset derived from metastatic melanoma patients prior to anti-PD-1 treatment | Hugo et al., 2016 | GEO: GSE78220 |
| Wnt pathway-focused Nanostring dataset derived from metastatic melanoma patients prior to anti-PD-1 treatment | This paper | GEO: GSE165745 |
| Nanostring data from ETC-159 treated patients | This paper | GEO: GSE167039 |
| Experimental models: Cell lines | ||
| BrafV600EPten−/− (BPD6, male) cell line | Hanks Lab | N/A |
| p53flox/flox;KrasG12D cell line | Hanks Lab | N/A |
| Lewis Lung Carcinoma | ATCC | ATCC Cat# CRL-1642; RRID:CVCL_4358 |
| DC2.4 | Kenneth Rock, University of Massachusetts Medical School | Shen et al., 1997; RRID: CVCL_J409 |
| HEK293-LEF1/TCF-luciferase cell line | N/A | Ring et al., 2011 |
| Experimental models: Organisms/strains | ||
| B6.CgBRAFtm1MmcmPTENtm1HwuTg(TyrCre/ERT2)13Bos/BosJ (BRAFV600E PTEN−/−, H-2b) transgenic mice | Jackson Labs | IMSR Cat# JAX:013590; RRID:IMSR_JAX:013590 |
| p53flox/flox;KrasG12D transgenic mice | Anton Berns, Netherlands Cancer Institute | DuPage et al., 2009 |
| C57BL/6 wild type | Jackson Labs | IMSR Cat# JAX_000664; RRID:IMSR_JAX:000664 |
| BALB/c (H-2d) wild type | Jackson Labs | IMSR Cat# JAX_000651; RRID:IMSR_JAX:000651 |
| B6.Cg-Foxp3tm2Tch/J (FoxP-EGFP, H-2b) | Jackson Labs | IMSR Cat# JAX:006772; RRID:IMSR_JAX:006772 |
| B6.CgBRAFtm1MmcmPTENtm1HwuTg(TyrCre/ERT2)13Bos/BosJ (BRAFV600E PTEN−/−, H-2b) transgenic mice | Jackson Labs | IMSR Cat# JAX:013590; RRID:IMSR_JAX:013590 |
| Oligonucleotides | ||
| Tcf7-forward: AGC TTT CTC CAC TCT ACG AAC A | IDT | N/A |
| Tcf7-reverse: AAT CCA GAG AGA TCG GGG GTC | IDT | N/A |
| Ccnd1-forward: GCG TAC CCT GAC ACC AAT CTC | IDT | N/A |
| Ccnd1-reverse: CTC CTC TTC GCA CTT CTG CTC | IDT | N/A |
| Gapdh-forward: GTC TAC ATG TTC CAG TAT GAC TCC | IDT | N/A |
| Gapdh-reverse: AGT GAG TTG TCA TAT TTC TCG TGG T | IDT | N/A |
| Cxcl2-forward: GAG CTT GAG TGT GAC GCC CCC AGG | IDT | N/A |
| Cxcl2-reverse: GTT AGC CTT GCC TTT GTT CAG TATC | IDT | N/A |
| Cxcl5-forward: GCA TTT CTG TTG CTG TTC ACG CTG | IDT | N/A |
| Cxcl5-reverse: CCT CCT TCT GGT TTT TCA GTT TAG C | IDT | N/A |
| Actb-forward: GTC TAC ATG TTC CAG TAT GAC TCC | IDT | N/A |
| Actb-reverse: AGT GAG TTG TCA TAT TTC TCG TGG T | IDT | N/A |
| Ccl4-forward: GCC CTC TCT CTC CTC TTG CT | IDT | N/A |
| Ccl4-reverse: GTC TGC CTC TTT TGG TCA GG | IDT | N/A |
| Software and algorithms | ||
| nSolver | Nanostring | RRID: SCR_003420 |
| Amira 3D Visualization and Analysis Software Suite | FEI, Thermo Fisher Scientific | RRID: SCR_007353 |
| ImageJ | ImageJ | RRID: SCR_003070 |
| FlowJo Version 10 | Tristar | RRID: SCR_008520 |
| Prism | GraphPad | RRID: SCR_002798 |
| ImmunoCapture and ImmunSpot | ImmunoSpot | RRID: SCR_011082 |
| R | R Project for Statistical Computing | RRID:SCR_001905 |
| Heatmap.2 | R Function | https://cran.r-project.org/web/packages/gplots/index.html |
| Other | ||
| XRAD225 Cx | Precision X-Ray, Inc | N/A |
| gentleMACS Tissue Dissociator | Miltenyi Biotec | Cat # 130-093-235 |
| FACSCanto II | Becton Dickinson | N/A |
| LSRII | Becton Dickinson | N/A |
| gentleMACS C-tubes | Miltenyi Biotec | Cat # 130-093-237 |
| gentleMACS M-tubes | Miltenyi Biotec | Cat # 130-093-236 |