Fig. 4

GATA3 was involved in METTL3-regulated transcription of SLC7A11. A The venn diagram showed the overlap of transcription factors of SLC7A11 predicted by PROMO, JASPAR, ChIPBase and ChIP-Atlas, respectively. B Heatmap of SLC7A11 predicted transcription factors. C The expression of GATA3 mRNA was checked by qRT-PCR in HeLa and MDA-MB-231 cells. D The expression of GATA3 protein was checked by western blot analysis in HeLa and MDA-MB-231 cells. E m⁶A RIP-qPCR analysis of GATA3 mRNA in HeLa WT and METTL3 KD cells. F The m⁶A in 5′UTR, CDS and 3′UTR of GATA3 in WT or METTL3 KD HeLa cells was analyzed by m⁶A-RIP-qPCR using fragmented RNA. G HeLa cells were transfected with siRNAs of GATA3, pGL3-SLC7A11-WT promoter and pRL-TK plasmid for 48 h, then the luciferase activity of F-Luc/R-Luc was detected using Dual-Lumi™ II Luciferase Assay Kit. H HeLa cells were transfected with siRNAs of GATA3 for 24 h, and SLC7A11 precursor mRNA expression was checked by qRT-PCR. I HeLa cells were transfected with siRNAs of GATA3 for 48 h, and GATA3 and SLC7A11 protein expression were checked by western blot analysis (left) and quantitatively analyzed (right). J HeLa cells were transfected with siRNAs of GATA3 for 24 h; they were co-transfected with pGL3-SLC7A11 promoter reporter and pRL-TK for another 24 h. The luciferase activity of F-Luc/R-Luc was detected using Dual-Lumi™ II Luciferase Assay Kit. K The expression of GATA3 mRNA was checked by qRT-PCR in HeLa and MDA-MB-231 cells after treating with erastin (5 µM) for 24 h. L The expression of GATA3 protein was checked by western blot analysis in HeLa and MDA-MB-231 cells after treating with erastin (5 µM) for 24 h. M HeLa cells were transfected with siRNAs of GATA3, and then treated with increasing concentrations of erastin for 48 h, and the relative cell viability was detected using Cell Counting Kit-8 kit. N Schematic representation of the mutated promoter in pGL3-Basic-SLC7A11 reporter. O HeLa cells were co-transfected with siRNAs of GATA3, pGL3-SLC7A11 mutant promoter and pRL-TK plasmid for 24 h, then the luciferase activity of F-Luc/R-Luc was detected using Dual-Lumi™ II Luciferase Assay Kit. P HeLa cells were co-transfected with pGL3-SLC7A11 mutant promoter reporter, pRL-TK and METTL3 plasmid for 24 h, and the luciferase activity of F-Luc/R-Luc was detected using 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