Fig. 5
YTHDF1-promoted YAP mRNA translation is regulated by m6A modification and interaction with eIF3a. a The mRNA and protein levels of YTHDF1 were analyzed by RT-PCR and western blot assays in paired fresh NSCLC tumor cancer tissues (Tumor) and matched adjacent normal tissues (Normal) (left panel, n = 10; right panel, n = 30). b The expression of YTHDF1 was analyzed by IHC assay in the different grades of lung cancer tissues. c High expression of YTHDF1 is associated with worse prognosis for NSCLC patients (P = 0.018). (d, e) A549 cells were transfected with indicated genes of YTHDF1. d The migration growth was analyzed by scratch assay. e The expressions of E-cadherin and Vimentin were analyzed by western blot assay. f The xenografted tumors (left panel) and the metastasizing lung tumors (right panel) originated from A549 cells with stable expression of indicated genes constructed by subcutaneous injection. g The protein levels of YAP, CTGF and Cyr61 were analyzed in A549 cells determined by WB assay. h Co-IPs performed using lysates collected from A549 cells with immunoprecipitation by either YTHDF1 or eIF3a antibodies. (i-k) The protein level of YAP was analyzed by ELISA assay in puromycin treated A549 cells with transfection with indicated genes. (l-p) A549 cell were transfected with indicated genes of YTHDF1 and YAP. l The protein levels of YTHDF1, YAP, CTGF and Cyr61 were analyzed by western blot assay. m The size and number of colons were analyzed by colony formation assay. n The cellular invasion and migration growth were analyzed by transwell assay. o The expressions of E-cadherin and Vimentin were analyzed by qPCR assay. p The relative of cleaved Caspas-3 was analyzed by western blot assay. Results were presented as mean ± SD of three independent experiments. *P < 0.05 or **P < 0.01 indicates a significant difference between the indicated groups. ns, not significant