RNA-seq in wild-type, METTL16-knockout, and METTL16 rescue NOMO-1 cells

METTL16 belongs to methyltransferase like (METTL) family and could install m6A marks on its substrates. Here, we uncover a tumor-promoting role of METTL16 in AML and LSC self-renewal. To explore the mechanism underlying the oncogenic function of METTL16 in AML, we performed m6A-seq and RNA-seq. Via integrated analysis of m6A-seq data and RNA-seq data, we identified two bona fide targets of METTL16, BCAT1 and BCAT2. METTL16 functions as an m6A methyltransferase to regulate expression of BCAT1 and BCAT2, which contribute to reprogramming BCAA metabolism. NOMO-1 cells overexpressing empty vector (EV) or METTL16 as indicated were followed with endogenous METTL16 knockout by viral transduction. EV is used as control for overexpression (OE). sgNS is used as control for sgRNAs. The positive-infected cells were selected with 2ug/ml puromycin for 1-2 weeks. The cells were collected and total RNA were isolated from the whole cells via Qiazol. Ribosome RNA was depleted from total RNA using Ribo-Zero plus rRNA depletion kit.