Fig. 5
YTHDF1 is recruited by METTL3 to enhance SLC7A11 mRNA stability and translation. A and B The knockdown efficiency after 48 h transfection of YTHDF1 siRNA and negative control siRNA (siNC) in PC9 cells was confirmed by RT-qPCR and western blot. C and D The relative mRNA expression determined by RT-qPCR and the protein expression showed by western blot of SLC7A11 after YTHDF1 knockdown in PC9 cells. E SLC7A11 mRNA half-lives (t1/2) showed by RNA decay rates followed by RT-qPCR after YTHDF1 knockdown in PC9 cells. Data were collected at indicated timepoints (0 h, 3 h, and 6 h) with actinomycin D (Act D, 5 μg/mL) treatment. F RIP-qPCR revealed the binding enrichment of YTHDF1 to SLC7A11 in METTL3 stable knockdown and negative control PC9 cells. G The protein levels of SLC7A11 showed by western blot in YTHDF1 knockdown H322 cells with METTL3 overexpression, compared with control H322 cells. H Flow cytometry analysis of intracellular ROS levels in YTHDF1 knockdown H322 cells with METTL3 overexpression, compared with control H322 cells. I Flow cytometry analysis for apoptotic cell proportion (Q2 + Q3) by Annexin V-FITC/PI staining in H322 cells. *P < 0.05, **P < 0.01, ***P < 0.001, ns, not significant