Fig. 2

ALKBH5-mediated m6A modification is involved in the low expression of CELF2. A The RIP qRT-PCR assay showed the enrichment of m6A-modified CELF2 in PC cells (BxPC-3 and MIA PaCa2) compared with the normal pancreatic cell line HPDE6-C7. B The positive correlation between the levels of ALKBH5 and CELF2 in pancreatic cancer tissue was analyzed by RT-PCR. C–F The effect of ALKBH5 silencing or overexpression on CELF2 expression. G The m6A modification locus 1116565 of CELF2 can be modified by ALKBH5 predicted by the online website http://m6avar.renlab.org/, which was confirmed by the following luciferase reporter assay. H and I RIP qRT-PCR results showing the enrichment of m6A-modified CELF2 after ALKBH5 silencing or overexpression. J CELF2 stability analysis in BxPC-3 cells with CELF2 silencing or overexpression in the presence of actinomycin D. *P < 0.05, **P < 0.01. K The enrichment of the YTHDF2 on CELF2 in PC cells was analyzed by RIP-RT-qPCR assay. L, M Effect of YTHDF2 knockdown or overexpression on CELF2 expression. N Actinomycin D assay results showed the effect of YTHDF2 knockdown or overexpression on CELF2 stability. O RIP-RT-qPCR assay results show the enrichment of YTHDF2 on CELF2 while ALKBH5 silencing. Data are shown as the mean SD of three replicates. *P < 0.05; **P < 0.01