Fig. 5.

YTHDC2 destabilized SLC7A11 mRNA in an m⁶A-dependent manner. (A, B) The decay rate of SLC7A11 mRNA in control cells, H1299 cells with YTHDC2^WT or YTHDC2^ΔYTH overexpression, and H1975 cells with or without YTHDC2 knocked out or reconstitution after ActD (5 μg/mL) treatment at the indicated times. (C) SLC7A11 protein and mRNA expression in H1299 control cells and YTHDC2^WT overexpression cells with or without EXOSC10 knocked out. (D) SLC7A11 protein and mRNA expression in H1299 control cells and YTHDC2^WT overexpression cells with or without DMSO, DEPC or RNasin (5 U/μl) treatment. (E–G) The m⁶A levels and decay rate of SLC7A11 mRNA were measured by m⁶A RNA methylation assay kit, m⁶A dot blot and RT-qPCR assays in control cells, H1975 cells with METTL3 knockdown, and H1299 cells with METTL3 overexpression. (H–J) The decay rate of SLC7A11 mRNA, cystine uptake and Lipid ROS generation were detected in H1299 control cells, YTHDC2 overexpression cells with or without knocked METTL3 down. Statistical analysis was performed using two-way ANOVA (A-D, G, H) and one-way ANOVA (E, I, J). Data are means ± SEMs, **p < 0.01, N·S.: no significant.