Fig. 5

METTL14 knockdown enhances SOX4 mRNA stability via an m6A-YTHDF2-dependent pathway. a. The m6A contents of total RNAs in METTL14-knockdown HCT116 and HCT8 cells (left panel) were detected using dot blot with m6A antibody. Methylene blue staining were served as the loading control (right panel). b. MeRIP-qPCR analysis was used to demonstrate METTL14-mediated SOX4 m6A modification in HCT116 and HCT8 cells. m6A modification of SOX4 was depleted upon METTL14 knockdown. c. Wild-type or m6A consensus sequence mutant SOX4 cDNA was fused with firely luciferse reporter. d. Mutation of m6A consensus sequences or knockdown of METTL14 relieved the posttranscriptional repression of SOX4 in HCT116 and HCT8 cells. e. The mRNA levels of YTHDF2 and SOX4 in YTHDF2 knockdown CRC cells were detected by qRT-PCR. f. The protein levels of YTHDF2 and SOX4 in YTHDF2 knockdown CRC cells were detected by western blot. GAPDH was used as control. g. Precursor and mature mRNA of SOX4 in METTL14 stable knockdown and control HCT116 cells. h. i. The precursor h and mature i SOX4 mRNA expression were detected at indicated times. j. RIP-qPCR assay using YTHDF2-specifc antibody and IgG control antibody to measure the enrichment of YTHDF2 binding to SOX4 m6A modification sites. *P < 0.05, **P < 0.01, ***P < 0.001