RNA m6A quantification and MeRIP-qPCR

The m6A modification level of total RNA was measured by EpiQuik m6A RNA Methylation Quantification Kit. The total RNA was bound in the detection hole with a solution with a high binding degree of RNA. m6A was detected by a specific capture m6A antibody. The absorbance was read at 450 nm, then the OD value was measured by colorimetry and the total m6A content was calculated. EZ-Magna RIP kit (Millipore) was used to evaluate the modification level of m6A in the specific gene according to the manufacturer’s instructions. In short, the RNA containing m6A methylated was coated with m6A antibody and immunoprecipitated on magnetic beads A/G. The enriched RNA-antibody complex was digested with protease to obtain m6A modified RNA. Then qRT-PCR was used to quantify the enriched RNA.