Fig. 6: RBM15 regulates the m6A modification of serine and glycine metabolic genes.
a The m6A content of total RNA in MDA-MB-231 and Hs578T BC cells with or without RBM15 silencing was determined. b The MeRIP-seq peaks of the indicated genes from the Gene Expression Omnibus were aligned. The methylated adenosine base is denoted with a red box and red “A”. c Methylated RNA in MDA-MB-231 and Hs578T cells. d MDA-MB-231 and Hs 578 T cells were transfected with siRBM15 or siCon and immunoprecipitated with a m6A antibody. Next, qRT‒PCR analysis was performed using primers against the PSAT1, PSPH, and SHMT2 mRNAs. e–g Diagram of the RBM15 mutant construct. e MDA-MB-231 and Hs578T cells were stably infected with SFB (S protein, Flag, and streptavidin-binding peptide)-tagged WT RBM15 or ΔRRM-RBM15. The infected cells were harvested, and protein expression levels were analyzed by WB using the indicated antibodies (f). Then, the infected cells were treated with DMSO or actinomycin D (Act D) and harvested at the indicated time points for qRT‒PCR with the indicated primers (g). The data are expressed as the means ± SDs of triplicate samples (*p < 0.05, **p < 0.01, ***p < 0.005, and ****p < 0.001).
