KEY RESOURCES TABLE
| REAGENT or RESOURCE | SOURCE | IDENTIFIER |
|---|---|---|
| Antibodies | ||
| Purified anti-β-actin (For WB) | Santa Cruz | Cat# sc-47778 |
| Purified anti-Vinculin (For WB) | Millipore | Cat# 05-386 |
| Purified anti-Mouse Irg1 (For WB and IF) | Cell Signalling Technology | Cat# 17805 |
| Purified anti-Human Acod1 (for IF) | Aviva Systems Biology | Cat# OACA09406 |
| Purified anti-C/EBPβ (For WB) | BioLegend | Cat# 606202 |
| Purified anti-Stat3 (For WB) | Cell Signalling Technology | Cat# 4904P |
| Purified anti-p-Stat3 (For WB) | Cell Signalling Technology | Cat# 9145 |
| Purified anti-Stat5 (D2O6Y) Rabbit mAb (For WB) | Cell Signalling Technology | Cat# 94205T |
| Purified anti-p-Stat5 (For WB) | Cell Signalling Technology | Cat# 9314S |
| Purified anti-Nrf2 (D1Z9C) (For WB) | Cell Signalling Technology | Cat# 12721T |
| Purified anti-Gpx4 Antibody (For WB) | R&D Systems | Cat# MAB5457-SP |
| Purified anti-Ly6G (Clone 1A8; For IF and neutrophil depletion) | BioXcell | Cat# BP0075-1 |
| Purified anti-CD11b (For IF) | BioLegend | Cat# 101202 |
| Purified anti-CD68 (for IF) | Bio-rad | Cat# 1602 |
| Purified anti-CD3 (Clone 145-2C11) (for IF) | BioLegend | Cat# 100302 |
| Purified anti-B220 (Clone RA3-6B2) (for IF) | BioLegend | Cat# 103201 |
| Purified anti-CD3ε [145-2C11] 500 μg (for T cell activation) | BioLegend | Cat# 100302 |
| Purified anti-CD28 (for T cell activation) | BioLegend | Cat# 102102 |
| AF488 anti-hCD15 | BioLegend | Cat# 301910 |
| Purified anti-Pan-Cytokeratin | BioLegend | Cat# 914204 |
| AF647 anti-Rabbit IgG (H+L) | Jackson ImmunoResearch | Cat# 711-605-152 |
| AF647 anti-Mouse IgG (H+L) | Jackson ImmunoResearch | Cat# 115-605-146 |
| AF647 anti-Rat IgG | Jackson ImmunoResearch | Cat# 712-606-153 |
| AF594 anti-Rabbit | Jackson ImmunoResearch | Cat# 611-585-215 |
| AF594 anti-mouse | Jackson ImmunoResearch | Cat# 615-585-214 |
| AF594 anti-Rat IgG | Jackson ImmunoResearch | Cat# 712-585-150 |
| HRP-conjugated anti-mouse IgG | Cell Signalling Technology | Cat# 7076P2 |
| HRP-conjugated anti-rabbit IgG | Cell Signalling Technology | Cat# 7074S |
| PE-Cy7 anti-mLY6G (1A8) | Tonbo Biosciences | Cat# 60-1276-U100 |
| APC anti-h/mCD11b (M1/70) | Tonbo Biosciences | Cat# 20-0112-U100 |
| APC anti-mCD45 (30-F11) | Tonbo Biosciences | Cat# 20-0451-U025 |
| PE-Cy7 anti-mCD45 (30-F11) | Tonbo Biosciences | Cat# 60-0451-U100 |
| PerCP-Cy5.5 anti-mCD45 (30-F11) | Tonbo Biosciences | Cat# 65-0451-U100 |
| PerCP-Cyanine5.5 anti-mCD3 (17A2) | Tonbo Biosciences | Cat# 65-0032-U100 |
| APC-Cy7 anti-mCD3 (17A2) | Tonbo Biosciences | Cat# 25-0032-U100 |
| PE anti-mCD4 (GK1.5) | Tonbo Biosciences | Cat# 50-0041-U100 |
| APC anti-mCD8a (53-6.7) | Tonbo Biosciences | Cat# 20-0081-U100 |
| PerCP-Cy5.5 anti-mCD8a (53-6.7) | Tonbo Biosciences | Cat# 65-0081-U100 |
| APC anti-mIFN gamma (XMG1.2) | Tonbo Biosciences | Cat# 20-7311-U100 |
| APC anti-mFoxp3 (3G3) | Tonbo Biosciences | Cat# 20-5773-U025 |
| Anti-mCD16 + CD32 (2.4G2) | Tonbo Biosciences | Cat# 70-0161-U500 |
| APC Anti-mCD274 (PD-L1) (10F.9G2) | Tonbo Biosciences | Cat# 20-1243-U025 |
| Purified anti-mGM-CSF (for blocking) | Tonbo Biosciences | Cat# 21-8339-U020 |
| Purified anti-Cleaved Caspase-3 (Asp175) Antibody | Cell Signaling Technology | Cat# 9661S |
| Lineage cocktail biotin antibody | Biolegend | Cat# 480050 |
| Percp5.5-Streptavidin | eBioscience | Cat# 45-4317-80 |
| APC anti-c-Kit | Tonbo Bioscience | Cat# 201173-U100 |
| PE anti-CD34 | Biolegend, | Cat# 128609 |
| APC-Cy7 anti-mouse CD16/32 (FcγRII/III) | Biolegend | Cat# 101327 |
| Pe-Cy7 anti-sca-1 | Biolegend | Cat# 122514 |
| Biological samples | ||
| FFPE sample of human breast cancer surgical specimen | Harper Cancer Research Institute at University of Notre Dame | N/A |
| Chemicals, peptides, and recombinant proteins | ||
| Collagenase D, from Clostridium histolyticum | Sigma-Aldrich | Cat# COLLD-RO |
| DNase I, Grade II, from bovine pancreas | Sigma-Aldrich | Cat# 10104159001 |
| RIPA Lysis and Extraction Buffer | Thermo Fisher | Cat# 89900 |
| Dapi, for nucleic acid staining | Sigma-Aldrich | Cat# D9542-10MG |
| Recombinant Mouse GM-CSF | BioLegend | Cat# 576302 |
| Recombinant Mouse M-CSF | BioLegend | Cat# 576402 |
| Recombinant Mouse G-CSF | BioLegend | Cat# 574602 |
| Recombinant Mouse CCL2 | BioLegend | Cat# 578402 |
| Recombinant Mouse CXCL2 | BioLegend | Cat# 582502 |
| Recombinant Mouse CCL5 | BioLegend | Cat# 594202 |
| Recombinant Mouse CXCL1 | BioLegend | Cat# 573702 |
| Recombinant Mouse CXCL10 | BioLegend | Cat# 573602 |
| TIMP-1 | BioLegend | Cat# 593702 |
| beta-Estradiol, suitable for cell culture | Sigma-Aldrich | Cat# E2758-250MG |
| STAT5-IN-1 | MedChem Express | Cat# HY-101853 |
| BP-1-102 | MedChem Express | Cat# HY-100493 |
| WP1066 | MedChem Express | Cat# HY-15312 |
| LLL12 | Sigma-Aldrich | Cat# 573131 |
| Dimethyl itaconate,99% | Sigma-Aldrich | Cat# 592498-25G |
| 4-octyl Itaconate | Cayman Chemical | Cat# 25374 |
| Itaconic acid,≥99% | Sigma-Aldrich | Cat# I29204-100G |
| Brefeldin A Solution (1,000X) | VWR | Cat# 420601-BL |
| PMA, for use in molecular biology applications, ≥99% (HPLC) | Sigma-Aldrich | Cat# P1585-25MG |
| Ionomycin, 98% | VWR | Cat# AAJ62448-MCR |
| C11 BODIPY 581/591 | Cayman Chemical | Cat# 27086 |
| APC Annexin V | Tonbo Biosciences | Cat# 20-6409-T100 |
| D-Luciferin, potassium salt | Goldbio | Cat# LUCK-4G |
| Ferrostatin-1 | Cayman Chemical | Cat# 17729 |
| ML385 | MedChem Express | Cat# HY-100523 |
| NK252 | MedChem Express | Cat# HY-19734 |
| Protease inhibitors | Bimake | Cat# B14012 |
| Phosphatase inhibitors | Roche | Cat# 4906845001 |
| Clarity Max™ Western ECL Substrate | Bio-Rad | Cat# 1705062 |
| Critical commercial assays | ||
| CellTrace™ CFSE Cell Proliferation Kit | Invitrogen | Cat# C34554 |
| MojoSort™ Mouse Ly-6G Selection Kit | Biolegend | Cat# 480124 |
| Mitochondrial ROS Detection Assay Kit | Cayman Chemical | Cat# 701600 |
| ROS Detection Cell-Based Assay Kit (DCFDA) | Cayman Chemical | Cat# 601520 |
| MojoSort™ Mouse CD3 T Cell Isolation Kit | Biolegend | Cat# 480024 |
| Foxp3 / Transcription Factor Staining Buffer Kit | Tonbo Biosciences | Cat# TNB-0607-KIT |
| Proteome Profiler Mouse Cytokine Array Kit, Panel A | R&D Systems | Cat# ARY006 |
| GM-CSF ELISA MAX™ Standard ELISA Kit | Biolegend | Cat# 432201 |
| Glutathione Assay Kit | Cayman Chemical | Cat# 703002 |
| BCA Protein Assay Kit | VWR | Cat# PI23225 |
| LEGENDplex™ Mouse Inflammation Panel | Biolegend | Cat# 740446 |
| Deposited data | ||
| Drop-seq raw data comparing mice Py7160 TIN and blood neutrophils | This paper | GEO: GSE216425 |
| Microarray data of human breast cancer metastases | PMID: 24287398 | GEO: GSE46141 |
| Single-cell sequencing of human TNBC metastatic tissue and peripheral blood | PMID: 34653365 | GEO: GSE169246 |
| RNA-seq data for murine breast cancer cells, tumor-infiltrating macrophages and tumor-infiltrating neutrophils | PMID: 31451770 | GEO: GSE104765 |
| PDF file containing uncropped scans of all Western blots | This paper | Data S1 |
| Excel file containing the values used to create all graphs | This paper | Data S1 |
| Experimental models: Cell lines | ||
| Py7160 | This study | N/A |
| E0771 | Siyuan Zhang lab at University of Notre Dame | N/A |
| E0771-TR | This study | N/A |
| 4T1 | Yibin Kang lab at Princeton University | N/A |
| 168FARN | Yibin Kang lab at Princeton University | N/A |
| Nmumg | Yibin Kang lab at Princeton University | N/A |
| ER-Hoxb8 progenitor cell line | David B. Sykes lab at Harvard University | N/A |
| SCF-producing CHO cell line | David B. Sykes lab at Harvard University | N/A |
| GMCSF-producing B16 cell line | David B. Sykes lab at Harvard University | N/A |
| Experimental models: Organisms/strains | ||
| Mouse: C57BL/6J | The Jackson Laboratory | RRID:IMSR_JAX:00 0664 |
| Mouse: FVB | The Jackson Laboratory | RRID:IMSR_JAX:00 1800 |
| Mouse: MMTV-PyMT | The Jackson Laboratory | RRID:IMSR_JAX:02 2974 |
| Mouse: Acod1−/− | The Jackson Laboratory | RRID:IMSR_JAX:02 9340 |
| Mouse: MRP8-Cre | The Jackson Laboratory | RRID:IMSR_JAX:02 1614 |
| Mouse: LysM-cre | The Jackson Laboratory | RRID:IMSR_JAX:00 4781 |
| Mouse: Acod1f/f | Michael Diamond lab from Washington University in St. Louis | N/A |
| Mouse: Rag1−/− | The Jackson Laboratory | RRID:IMSR_JAX:00 2216 |
| Mouse: Ly6G-cre | Matthias Gunzer and Anja Hasenberg lab from University Hospital Essen | N/A |
| Oligonucleotides | ||
| qPCR primers | This paper | Table S5 |
| Hashtag antibody barcode sequence | Biolegend | Table S5 |
| Recombinant DNA | ||
| Csf2 sgRNA CRISPR/Cas9 All-in-One Lentivector set (Mouse) | abm | Cat# 16914114 |
| CRISPR Scrambled sgRNA All-in-One Lentiviral Vector (with spCas9) | abm | Cat# K010 |
| Three MISSION lentiviral shRNA constructions in the pLKO backbone targeting mouse Cebpb | Sigma-Aldrich | Cat#TRCN0000231408, TRCN0000231410 and TRCN0000231411) |
| pLKO vector with scramble shRNA | Sigma-Aldrich | Cat# SHC016-1EA |
| psPAX2 | Addgene | Cat# 12260 |
| pMD2.G | Addgene | Cat# 12259 |
| Software and algorithms | ||
| R studio | Posit | https://www.rstudio.com/products/rstudio/ |
| Graphpad Prism 8.0 software | GraphPad Software, Inc. | https://www.graphpad.com |
| FiJi software | https://imagej.net/Fiji/Downloads | RRID:SCR_002285 |
| FlowJo | FlowJo LLC | https://www.flowjo.com |
| Seurat R package v4.0 | Satija Lab | https://satijalab.org/seurat/ |
| Ingenuity Pathway Analysis (IPA) | Qiagen | https://digitalinsights.qiagen.com/products |
| Auro Imaging Software | Spectral Instruments Imaging, LLC | https://spectralinvivo.com/software/ |