Fig. 4
circPDE5A decreases the m6A modification of EIF3C mRNA and restrains its translation. A, m6A motifs identified using MeRIP-seq. B, The distribution of m6A modification in 3′UTR, 5′UTR, and exon regions after circPDE5A overexpression or knockdown. C, Venn diagram showing the overlapped altered m6A modification genes after circPDE5A overexpression or knockdown. D, Integrative genomics viewer displaying the results of EIF3C m6A modification distribution in MeRIP-seq. E, F, Left: MeRIP assay showing the m6A modification level of EIF3C in 22Rv-1 cells with circPDE5A knockdown (E) or overexpression (F); right: the mRNA and protein expression of EIF3C in 22Rv-1 cells with circPDE5A knockdown (E) or overexpression (F). G, RT-qPCR assay showing the cellular distribution of EIF3C in 22Rv-1 cells with circPDE5A overexpression. H, RT-qPCR assay showing the EIF3C mRNA stability in 22Rv-1 cells with circPDE5A overexpression. I, WB assay showing the EIF3C protein stability in 22Rv-1 cells with circPDE5A overexpression or knockdown. J, RT-qPCR assay showing the relative level of EIF3C mRNA in gradient fractions with circPDE5A overexpression in C4-2B and 22Rv-1 cells. Data represents mean ± S.D. from three independent experiments. ***, p < 0.001