Fig. 6

PA2G4 increases FYN expression in a YTHDF2-dependent manner. A MeRIP assay for detecting the m6A modification on the mRNA of FYN. Data was shown as percentage of FYN mRNA in anti-m6A precipitates against 10% input. B HCC cells were transfected with siRNA targeting YTHDF2 (B1) or pcDNA3.1-YTHDF2 (B2). qRT-PCR analyses were performed to evaluating the mRNA level of YTHDF2 and FYN 72 h after transfection. Data normalized to GAPDH and are shown as fold change to their respective control cells. C HCC cells were treated as in B, Western blot analyses of the indicated proteins. GAPDH as loading control. D HCC cells were transfected with siRNA targeting YTHDF2 for 72 h before administration of actinomycin D. qRT-PCR was performed to determine the relative FYN mRNA expression after treating with actinomycin D for the indicated time in each group. Data normalized to HPRT1 and are shown as percentage of the relative level in 0 h. E Huh7 cells with PA2G4 knocked down were transfected with siYTHDF2-1#, pcDNA3.1-YTHDF2 and pcDNA3.1-YTHDF2-5 A for 72 h. Western blot analysis of the indicated proteins. GAPDH as loading control. F Cells were treated as in E, tranwell assays were then performed to evaluate the cell migration and invasion abilities. Quantification of cells migrated and invaded. Data was shown as Mean ± SD. Two-tailed Student t test for (A, B, D and F). *p < 0.05, **p < 0.01. si: small interference; NC: negative control