Fig. 7
RBM15 induces m6A RNA methylation of R-loops. A Validation of S9.6 IP LC-MS/MS results by Western blot assay. Experiments were performed with HA-purified RBM15 protein and S9.6 antibody in DU-145 and PC-3 cells. B CRISPR-Cas9 mediated KO of RBM15 in PC-3 cells as detected by western blot. C PC-3 cells were immunostained for anti-RBM15 and S9.6; (Left): Representative images are shown (scale bar: 60 μm). (Right): Quantification of cellular immunofluorescence (n = 10 per group). Data are presented as means ± 95CI, two-tailed unpaired t-test. D The R-loop levels were assessed in RBM15 overexpression cells and RBM15 KO cells compared to control. F RBM15 KO DU-145 and PC-3 cells were used to evaluate total RNA and chromatin-associated RNA m6A levels by dot-blot. G MeDIP-qPCR analysis indicated the m6A site in R-loops at SEMA3F promoter, and the m6A levels could be upregulated by RBM15 overexpression, Data are presented as means ± SD, two-tailed unpaired t-test. H DU-145 and PC-3 cells were transfected with pc-RBM15 and pcDNA 3.1 vector. The total RNA and chromatin-associated RNA m6A levels were evaluated by dot-blot. *p-value < 0.05, **p-value < 0.01, ***p-value < 0.001, ****p-value < 0.0001