Extended Data Fig. 4: Cancer cell-intrinsic effect of inflammatory ccRCC cells on neutrophil phenotypes.
From: Epigenetic remodelling shapes inflammatory renal cancer and neutrophil-dependent metastasis
a, Transwell assay of BM-derived neutrophils from intact mice. Neutrophils were allowed to migrate for 2 h toward culture supernatants from the indicated cells. n = 3 mice/group. One-way ANOVA and Tukey’s test. b, Apoptosis-detection assay for BM-derived neutrophils from intact mice. Neutrophils were co-cultured with culture supernatants for 1 d. (left) Representative panel for detecting apoptotic cells. The percent of each fraction is indicated. (right) Percentage of Annexin V+ PI– cell population. n = 3 mice/group. One-way ANOVA and Tukey’s test. c, RNA-sequencing analysis of peripheral neutrophils. Heat map showing expression of the top 10 genes increased in neutrophils from OS-RC-2 derivative- compared to vehicle- and OSPa cell-inoculated mice. n = 2 mice/group. Genes were screened following the criteria; RPKM ≥1 in any of neutrophils. d, e, qRT-PCR analysis for the indicated mRNAs in BM-derived neutrophils from OS5K-3-bearing (d) or 786-3K-bearing (e) mice, co-cultured with vehicle or culture supernatants of OS5K-3 (d) or 786-3K (e) cells, respectively. Log2-transformed fold-changes compared to vehicle were indicated. n = 3 (d; Ms4a4a), n = 7 (d; Ms4a6c and Ms4a6d), and n = 4 (e) mice. Two-sided Welch’s t test. f, qRT-PCR analysis for the Ms4a6d mRNAs in neutrophils derived from bone, blood, and tumour of mice bearing OSPa or OS5K-3 cells. n = 2 mice/group. In all panels in this figure, the bars represent means ± S.D.
