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  • STAR METHODS br Detailed methods are

    2020-08-30

    STAR+METHODS
    Detailed methods are provided in the online version of this paper and include the following:
    d KEY RESOURCES TABLE
    d CONTACT FOR REAGENT AND RESOURCE SHARING d EXPERIMENTAL MODELS
    B Mice
    B Bone Marrow Transplantation
    (G) Representative confocal images of spleen-derived neutrophils from tumor-bearing mice (‘‘green‘‘ Ly6G) ingesting zymosan beads (‘‘red‘‘) counterstained with DAPI.
    (H) Quantification of neutrophils with ingested Salmonella, N R 10.
    (I) Quantification of zymosan beads number ingested per neutrophil isolated from indicated mice, N R 10.
    (J) qRT-PCR analysis of the expression of indicated antibacterial peptides in tumors lysates of Ly6GCre+Il1rf/f or control BM transplanted mice, NR8. Data are means ± SEM. Representative of at least 2 independent experiments. See also Figure S7.
    B Antibiotic treatment
    B Immune Glycoursodeoxycholic acid isolation, ex vivo cell stimulation, Flow Cytometry and Cell Sorting
    B Intestinal epithelial cell isolation for protein analysis B Nuclear protein extraction and western blot
    B In Vivo Treatment with Anakinra
    B RNA extraction and quantitative Real Time-PCR Analysis
    B Analysis of Cytokine Production by Multiplex or ELISA B Immunohistology and immunofluorescence
    B Whole tissue IF staining for bacteria with Yo-Yo 1 dye B Neutrophils or macrophage isolation from bone
    marrow, spleen, peritoneal cavities and tumors B Bacteria killing assay
    B Phagocytosis assay B Microbiome analyses
    d QUANTIFICATION AND STATISTICAL ANALYSIS d DATA AND SOFTWARE AVAILABILITY
    SUPPLEMENTAL INFORMATION
    Supplemental Information includes seven figures and can be found with this article online at https://doi.org/10.1016/j.immuni.2018.11.015.
    ACKNOWLEDGMENTS
    We thank S. Balachandran and G. Rall (FCCC), D. Mucida (Rockefeller Univer-sity) and R. Locke for critical comments, Dr. E. Koltsova (FCCC) for the help with confocal imaging and reading the manuscript, Aliia Fatkhullina (FCCC) for the help with graphical abstract, and E. Fearon (U Michigan) for CDX2Cre, CDX2ERT, and Apcf/f mice. We thank Histology laboratory of Department of Pathology at FCCC for tissue processing and slide preparation and LAF, Flow Cytometry, Cell Culture and Microscopy Facilities at FCCC. This work was supported by NIH P30 CA-006927 to FCCC, DFG grant CRC/TR 128 TPA03 and TPA07 to A.W., RSF Grant N14-15-00894 to M.R.K.; as well as by NIH R00 DK088589 and R01CA227629, Pew Scholar in Biomedical Sci-ences and AACR-Landon Innovator Awards, ACS-IRG #15-175-22, PA DOH CURE and US-Israel BSF grants to S.I.G. This project has been funded in whole or in part with federal funds from the Frederick National Laboratory for Cancer Research, National Institutes of Health, under contract HHSN261200800001E. The content of this publication does not necessarily reflect the views or policies of the Department of Health and Human Services, nor does mention of trade names, commercial products, or organizations imply endorsement by the US Government.
    AUTHOR CONTRIBUTIONS
    O.D.-P. performed experiments, analyzed the data, and wrote the manuscript; D.F.P. and V.H. provided technical assistance with animal work and reviewed and edited the manuscript; A.D., W.Y., V.T., and G.T. performed sequencing and analysis of microbiome and reviewed and edited the manuscript; I.A.M., A.W. and M.G. generated Il1r1f/f and Ly6GCre-Il1r1f/f mice, discussed the data and reviewed and edited the manuscript; I.P.S. and M.R.K. discussed the data and reviewed the manuscript; S.I.G. designed the study, analyzed and interpreted data, obtained funding, and wrote the manuscript with the input from all of the co-authors.
    DECLARATION OF INTERESTS
    The authors declare no competing interests.
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