Figure 4: Methylation of the miR-145 Promoter Was Mediated by DNMT1 and Tet2 Dynamic Alteration

(AandB) DNMT1 knockdown or Tet2 overexpression efficiency and the effect on miR-145 expression in vascular smooth muscle cells treated with TNF-α. Values are mean ± SEM of 3 samples in each group; *p < 0.05. (C) Methylation-specific polymerase chain reaction of the +129 CpG site in vascular smooth muscle cells treated with TNF-α with knockdown of DNMT1 or overexpression of Tet2. The methylated specific bands were obvious in the sh-con or Plenti-con groups treated with TNF-α, whereas the unmethylated specific bands were obvious in the sh-DNMT1 + TNF-α group and Plenti-Tet2 + TNF-α group. (D) Chromatin immunoprecipitation assay identified the miR-145 promoter sequence binding with DNMT1 and Tet2. (E) The relative enrichment rate of the chromatin immunoprecipitation assay was determined by real time polymerase chain reaction. Values are mean ± SEM of 3 samples in each group; *p < 0.05 vs. the con group. IgG = immunoglobulin G; mRNA = messenger ribonucleic acid; sh- = small hairpin RNA targeting; other abbreviations as in Figures 2 and 3.

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