Fig. 2
METTL3 triggered ferroptosis via suppressing SLC7A11 transcription. A The volcano plot showed the mutated genes between WT and METTL3 KD cells. B The overlap between the genes involved in ferroptosis and the mutated genes in METTL3 KD cells. C, D The mRNA expression of SLC7A11, ALOX5 and GLCM in HeLa (C) and MDA-MB-231 (D) was checked by qRT-PCR. E The protein expression of SLC7A11 in HeLa and MDA-MB-231 cells was checked by western blot analysis. F HeLa and MDA-MB-231 cells were transfected with vector control, METTL3 plasmid, and SLC7A11 plasmid for 24 h, and lipid ROS production was assayed by flow cytometry using C11-BODIPY. G, H HeLa (G) and MDA-MB-231 (H) HeLa cells were transfected with vector control, METTL3 plasmid, and SLC7A11 plasmid for 24 h, and the relative cell viability was detected using Cell Counting Kit-8 kit after treatment with erastin for 48 h. I m6A RIP-qPCR analysis of SLC7A11 mRNA in HeLa and MDA-MB-231 cells. J After treatment with Act-D for the indicated times, the mature mRNA levels of SLC7A11 were checked in WT and METTL3 KD HeLa cells by qRT-PCR. K WT or METTL3 KD HeLa cells were treated with 10 μg/ml CHX for the indicated time periods, and then the protein expression of SLC7A11 was detected by western blot (left) and quantitatively analyzed (right). L Polysome profiling of WT or METTL3 KD HeLa cells was analyzed. M Analysis of SLC7A11 mRNA in polysome/total for the METTL3 KD cells compared to control cells. N, O The SLC7A11 precursor mRNA expression HeLa and MDA-MB-231cells was checked by qRT-PCR. P, Q The luciferase activity of the SLC7A11 promoter in the F-Luc/R-Luc system was measured using the Dual-Lumi™ II Luciferase Assay Kit. 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