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. Author manuscript; available in PMC: 2019 Sep 1.
Published in final edited form as: Nat Cell Biol. 2018 Aug 27;20(9):1084–1097. doi: 10.1038/s41556-018-0173-5

Figure 4. Myeloid cell-derived IL-1β; is sufficient to prevent MIC differentiation.

Figure 4.

a, Top 10 differentially up-regulated genes (RNA-seq) in lungs from Met1 primary tumor bearing FVB mice relative to control lungs at day 14 (n=4 mice/cohort). b,qPCR on lungs 14 days following tail vein injection of mMICs into FVB mice bearing Met1 primary tumors or PBS control (per Fig. 1a, b). IL-1β; normalized relative to β-actin; (n=3 lungs/cohort, triplicate). c, Flow cytometric analysis of intracellular IL-1β in CD45+ cells from subset of lungs of Nude mice bearing Met1 primary tumors (n=3 animals) or PBS control (n=5 animals). d,qPCR on hMIC tumors at d28 using indicated human- and mouse-specific primers. n=3 tumors/cohort analyzed in triplicate.e, Flow cytometric analysis of intracellular IL-1β in CD11b+F4/80+ cells harvested from hMIC tumors grown opposite Matrigel control (n=8 tumors) or HMLER primary tumors (n=7 tumors); per Supplementary Fig. 2e. f, Schematic showing single injection xenograft model; applies to (g-i). g, Merged immunofluorescence images of hMIC tumors injected in Matrigel containing PBS (hMIC+PBS) or 10pg/ml IL-1β (hMIC + IL-1β) representing 2 independent experiments. Block arrows: examples of ZEB1+ tumor cells; long arrows: examples of ZEB1-tumour cells; arrowheads: ZEB1+ stromal cells). h, ECAD+LgT+ cells (hMIC+PBS n=10 independent images representing 7 tumors; hMIC+IL-1β n=9 independent images representing 6 tumors) or ZEB1+LgT+ cells (hMIC+PBS n=15 independent images representing 5 tumors; hMIC+IL-1β n=21 independent images representing 7 tumors) as % total number LgT+ tumor cells/microscopic field. i,Representative tumors stained with Mac2 (macrophages, brown) or hematoxylin (nuclei, blue), representing 2 independent experiments. All scale bars = 100 μm. Source data for b, c, d, e, h provided in Supplementary Table 1. 1-sided t test (b); 2-sided Mann- Whitney (c, h); 2-sided Welch’s t test (e); 2-way ANOVA followed by Sidak’s multiple comparison test (d).