Fig. 4

YTHDF3 recruits eIF4G2 complex to driver circ-YAP translation. A. Western blot testing the levels of the indicated proteins in circ-YAP-overexpressing 293T cells after knockdown of YTHDF1/2/3. B, C. Western blot testing the effect of YTHDF3 knockout on YAP-220 level. D. The indicated vectors were transfected into LoVo and SW620 cells, followed by western blot analysis of YAP-220 level. E. RIP assay using anti-YTHDF3 antibody, followed by qRT-PCR analysis of circ-YAP enrichment. F, G. The in vivo and in vitro pull-down assays using biotin-labeled circ-YAP probes, followed by western blot analysis of YTHDF3 level. H. Co-IP assay testing the interaction between YTHDF3 and eIF4G2 proteins. I. Western blot testing the effect of eIF4G2 silencing on YAP-220aa expression. J, K. RNA pull-down assays using biotin-labeled circ-YAP probes in YTHDF3^−/− SW620 and LoVo cells, followed by western blot analysis of the indicated protein levels. L, M. Transwell assay testing cell invasion in circ-YAP-overexpressing SW480 and HT29 cells after YTHDF3 knockout or eIF4G2 silencing. *P < 0.05, ***P < 0.001. Data (E, L, M) are the mean ± SD of three independent experiments carried out in triplicate. The uncropped western blot data are provided as a Original Blot Image file