Fig. 5
The p53 3' UTR mediated hnRNPA2B1 regulation of pre-mRNA stability. A MeRIP-seq data in HCT116 cells showed the distribution of m6A peaks along p53 mRNA. One significant m6A peak (chr17:7,571,893–7,572,013) was distributed in the 3'UTR region of p53 mRNA. B-C IB analysis of Flag-p53 levels in the HCT116 cells expressing Flag-p53 with the p53 CDS-3'UTR or p53 CDS and treated with control or hnRNPA2B1 siRNAs. D-E qRT-PCR analysis of Flag-p53 levels in the HCT116 cells expressing Flag-p53 with the p53 CDS-3'UTR or p53 CDS and treated with control or hnRNPA2B1 siRNAs. The sequence of the forward primer corresponds to the 3 × Flag vector. F IB analysis of Mettl3 levels in the HCT116 cells transfected with Mettl3 siRNAs. G qRT-PCR analysis of the expression of p53 pre-mRNA in the HCT116 cells transfected with Mettl3 siRNAs compared to those transfected with the siRNA control. H IB analysis of Mettl3 and hnRNPA2B1 levels in the LoVo cells transfected with Mettl3 plasmid. I qRT-PCR analysis of the expression of p53 pre-mRNA in the LoVo cells transfected with Mettl3 plasmid compared to those transfected with the vector. J-K MeRIP-qPCR analysis of m6A enrichment of p53 3'UTR in the CRC cells transfected with Mettl3 siRNA or overexpression plasmid compared to the cells transfected with the siRNA control or empty vector. L-M RIP analysis of the interaction of hnRNPA2B1 with p53 pre-mRNA in the CRC cells transfected with Mettl3 siRNA or overexpression plasmid compared to the cells transfected with the siRNA control or empty vector. N–O Relative activity of the wild-type or mutant p53 3'UTR firefly luciferase reporter in the HCT116 cells cotransfected with control or hnRNPA2B1 siRNAs and empty vector or Mettl3 overexpression plasmid. Data are shown as the mean ± SD of three independent experiments (*, P < 0.05; **, P < 0.01)