Figure 2: Methylation Status of the miR-145 Promoter In Vivo and In Vitro

(A) The CpG island of the miR-145 promoter was predicted, and primers were designed using MethPrimer (23). (B and C) BSP and the relative methylation ratio (20 clones) of each CpG site of the miR-145 promoter in aortas of ApoE−/− mice fed a Western diet (0, 12, and 24 weeks) are shown. Values are mean ± SEM from 6 animals in each group; *p < 0.05. (D) Level of miR-145 in vascular smooth muscle cells treated with TNF-α (10 ng/ml) or 5-aza (0, 2.5, or 5 nmol). Values are mean ± SEM of 3 samples in each group; *p < 0.05. (E and F) BSP and methylation rate of the miR-145 promoter in vascular smooth muscle cells treated with TNF-α or 5-aza. Values are mean ± SEM of 3 samples in each group; *p < 0.05, ***p < 0.01. 5-aza = 5-aza-2ʹ-deoxycytidine; BSP = bisulfite genomic sequencing; Con = control; TNF = tumor necrosis factor.

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