Methylated RNA immunoprecipitation sequencing in the Kasumi-1 cell line transfected with WTAP shRNA or scramble control shRNA

To evaluate the role of WTAP in modulating the m6A abundance, we conducted m6A sequencing in the Kasumi-1 cell line transfected with a lentivirus-mediated short hairpin RNA (shRNA) targeting WTAP gene or a scramble control shRNA. Stable silencing of the WTAP gene in leukemia cells was achieved through a lentiviral vector-based system for RNAi (GV112/hU6-MCS-CMV-Puromycin) constructed by GeneChem Technologies (China). Transfection of lentivirus was conducted according to the manufacturer's instruction. Kasumi-1 cells were seeded into 6-well plates overnight before transfection and the medium was changed approximately 12 to 16 hours after transfection. Forty-eight hours after shRNA transfection, cells were selected with puromycin (2 microgram per milliliter) for at least 24 hours before further investigations. A total of 3 biological replicates were performed.