The METTL3:METTL14 RNA methyltransferase regulates gene expression in MDA-MB-231 breast cancer cells

siRNAs targeting METTL3 and METTL14 were used to deplete METTL3 and METTL14 in MDA-MB-231 breast cancer cells and the effects on gene expression determined. Overall design: MDA-MB-231 cells were maintained in in RPMI-1640 medium supplemented with 100 µg/mL penicillin, 100 µg/mL streptomycin, 1 mM sodium pyruvate and 10% fetal bovine serum (FBS). siRNA-mediated functional depletion of METTL3 and METTL14 was performed in MDA-MB-231 breast cancer cells grown in phenol red-free RPMI-1640 medium supplemented with 1% penicillin-streptomycin, 2mM L-Glutamine and 1 mM sodium pyruvate and 10% FBS. ON-TARGETplus non-targeting control siRNAs (#D-001810-10-05, GE DharmaconTM) were employed as negative controls. Cells were transfected using DharmaFECT Transfection Reagent (GE DharmaconTM) and transfection was performed accoding to the manufacturer's instructions.The effect on global gene expression and alternative splicing was determined by RNA sequencing.