Fig. 7

YTHDF2 participates in the m6A-mediated regulation of ITGB4. a, b protein levels of indicated genes in si-YTHDF2 (a), si-IGF2BP2 (b) and si-NC treated ACHN cells detected by western blot. c Protein expressions of YTHDF2 and ITGB4 in Caki-1 cells with or without YTHDF2 overexpression detected by western blot. d Alterations of ITGB4 mRNA expression in YTHDF2-depletion ACHN cells and YTHDF2-overexpression Caki-1 cells respectively observed by qRT-PCR assay, respectively. e, f after treatment of cells with transcription inhibitor (TI) actinomycin D at 5 μg/ml for indicated times, ITGB4 mRNA expression in the cells with indicated transfections detected by qRT-PCR. g Enrichment of ITGB4 mRNA interacted with YTHDF2 in ACHN cells and Caki-1 cells detected by RIP assay with use of YTHDF2-specific antibody and IgG antibody as negative control. h, i RIP assay performed to detect alterations of the interaction between YTHDF2 and ITGB4 mRNA after METTL14 was depleted in ACHN cells (h) and overexpressed in Caki-1 cells (i). j Expressions of indicated proteins in Caki-1 cells with indicated treatments detected by western blot. An independent triplicate was conducted for each experiment. Bar graphs: means ± SDs. *p < 0.05, **p < 0.01 and ***p < 0.001. ns, not significant