Extended Data Fig. 8: Effect of the BETi on inflammatory ccRCC cells.
From: Epigenetic remodelling shapes inflammatory renal cancer and neutrophil-dependent metastasis
a, qRT-PCR analysis of the indicated mRNAs in BM-derived neutrophils. Neutrophils were co-cultured with culture supernatants from JQ1-pretreated OS5K-3 cells for 1 d. Log2-transformed fold-changes compared to those from DMSO-pretreated OS5K-3 cells were indicated. n = 3 mice. Two-sided Welch’s t test. b, Transwell assay of BM-derived neutrophils from intact mice. Neutrophils were incubated for 2 h and allowed to migrate toward the culture supernatants of JQ1-pretreated 786-3K cells. n = 3 mice. Two-sided Student’s t test. c, Apoptosis-detection assay of BM-derived neutrophils from intact mice. Neutrophils were co-cultured with culture supernatants of JQ1-pretreated 786-3K cells for 1 d. (left) Representative panel showing the detection of apoptotic cells. The percent of each fraction is indicated. (right) Percentages of Annexin V+ PI– cell populations. n = 3 mice. Two-sided Student’s t test. d, Apoptosis assay of BM-derived neutrophils. Neutrophils were co-cultured with culture supernatants of OS5K-3 cells containing SB265610 for 1 d. (left) Representative panel showing the approach used for detecting apoptotic cells. The percentages of each fraction are indicated. (right) The percentages of Annexin V+ PI– cell populations. n = 3 mice. Two-sided Student’s t test. e, Heat map showing expression of the indicated genes. RNA-expression data were from the RNA-sequencing analysis shown in Figs. 3–5, and Extended Data Fig. 5. f, Apoptosis-detection assay of BM-derived neutrophils from intact mice. Neutrophils were co-cultured with culture supernatants of OS5K-3 cells containing the indicated neutralizing antibodies for 1 d. (left) Representative panel showing the detection of apoptotic cells. The percent of each fraction is indicated. (right) Percentages of Annexin V+ PI– cell populations. n = 3 mice. One-way ANOVA and Tukey’s test. g, qRT-PCR analysis of the indicated mRNAs in OS5K-3 cells. The cells were treated with JQ1 for 1 d. Log2-transformed fold-changes compared to DMSO-treated OS5K-3 cells were indicated. n = 3 biologically independent samples.
