Fig. 1

METTL3 facilitated erastin-induced ferroptosis and enhanced mitochondrial ROS. A Cells were treated with or without 2 μM erastin for 24 h, and the m⁶A/A ratios of mRNA were checked by HPLC/MS/MS. B Dot blot (left) and quantitative analysis (right) were used to detect the total mRNA m⁶A. C HeLa cells were treated with erastin (2 μM), ferrostatin-1 (1 μM), acetylcysteine (NAC, 2 mM), Z-VAD-FMK (10 μM) and necrosulfonamide (NSA, 5 μM) for 24 h, and the m⁶A/A ratios of mRNA were checked by HPLC/MS/MS. D, E HeLa (D) and MDA-MB-231 (E) cells were treated with 2 μM erastin for 24 h, and the protein expression was assessed by western blot (left) and quantitative analysis (right). F, G HeLa (F) and MDA-MB-231 (G) cells were treated with increasing concentrations erastin for 48 h, and the relative cell viability was detected using Cell Counting Kit-8 kit. H HeLa cells were treated with or without erastin (0.1 μM) for 14 days, and the colonization capability was evaluated (left) and analyzed (right). I Representative transmission electron microscopy images of WT or METTL3 KD HeLa cells treated with erastin (2 μM, 24 h) (up). Yellow arrow represent shrunken mitochondria. The length of mitochondria was measured by image J (down). J WT and METTL3 KD HeLa cells were treated with or without erastin (2 μM) for 24 h, and lipid ROS production was assayed by flow cytometry using C11-BODIPY. K HeLa cells were transfected with vector control, METTL3 WT plasmid, or METTL3 DA mutant plasmid for 24 h, and lipid ROS production was assayed by flow cytometry using C11-BODIPY. Data are presented as mean ± SD from three independent experiments. *p < 0.05, **p < 0.01, ***p < 0.001, ns, no significant, by Student’s t test between two groups and by one-way ANOVA followed by Bonferroni test for multiple comparison