Fig. 3: HG and Nr-CWS regulate METTL3 transcription in HUVECs through H3K27ac mediated by KLF9.

A Data from the UCSC genome bioinformatics website (http://genome.ucsc.edu/) showed high enrichment of H3K27ac in the promoter of Mettl3. B ChIP-Qpcr analysis identified enrichment of H3K27ac in the promoter of Mettl3 in HUVEC cells. C Examination of the expression levels of the H3K27ac protein in HUVECs across different environments (n = 3 per group). D Venn diagram displays Mettl3-regulated genes from the tftarget and gencards database. And QPCR validation in different environments. E Protein levels of KLF9, H3K27ac, and METTL3 in different environments (n = 3 per group). F KLF9, H3K27ac, and METTL3 protein levels under different Nr-CWS treatment times (n = 3 per group). G After silencing KLF9 using small interfering RNA in the MM environment, the protein content of KLF9, H3K27ac, and METTL3 changed (n = 3 per group). H After overexpression of KLF9, the protein content of KLF9, H3K27ac, and METTL3 changed (n = 3 per group). I Schematic representation of KLF9 binding sites and Mettl3 promoter region with transcription start site (TSS). J Sequence logo representing the consensus Klf9 binding motif. K ChIP-qPCR validation of Klf9 binding at two sites within the METTL3 promoter, with siRNA-mediated Klf9 knockdown. L Luciferase reporter assay results showing relative promoter activity of wild-type and mutant Mettl3 promoters. M Western blot analysis demonstrating the effect of Klf9 knockdown on H3K27ac and METTL3 expression (n = 3 per group). N Graphical summary illustrating the proposed mechanism where KLF9 regulates H3K27ac and METTL3 in high-glucose and Nr-CWS conditions. The data are represented as mean ± SD of at least three independent experiments. *p < 0.05, **p < 0.01, ***p < 0.001 and ****p < 0.0001 are considered significant. H3K27ac H3K27 acetylation; ChIP chromatin immunoprecipitation.