Fig. 3.

FTO regulates the level of m⁶A modification in ESCC cells. A The content of m⁶A in total mRNA in five ESCC cell lines and normal esophageal epithelial cell (HEEC) was determined by the m⁶A RNA methylation quantitative kit. B, C The RNA m⁶A content was determined with the m⁶A methylation kit after overexpression FTO (FTO-OE) in HEEC (B) and FTO-knockdown (sh-FTO) in KYSE150 cells (C). D, E Dot blot assay was used to analyze the methylation of m⁶A RNA after overexpression FTO (FTO-OE) in HEEC (D) and FTO-knockdown (sh-FTO) in KYSE150 cells (E). F Predominant consensus motif GGAC was detected in both the control and sh-FTO KYSE150 cells in m⁶A-seq. G Proportion of m⁶A peak distribution in the exon, intergenic, intron, stop-codon, transcription start site (TSS), 3′-untranslated region (3′UTR), 5′-untranslated region (5′UTR) across the entire set of mRNA transcripts. H Density distribution of m⁶A peaks across mRNA transcripts. Regions of the 5′UTR, coding region (CDS), and 3′UTR were split into 100 segments, the percentage of m⁶A peaks in each segment was determined. The data was presented as the mean ± SDs (n = 3). The two-tailed Student’s t-test and one-way ANOVA were used to perform comparison between two groups and more groups, respectively. *p < 0.05,**p < 0.01; ****p < 0.0001