Figure 3: Expression and Activity of Methylation-Associated Proteins In Vivo and In Vitro

(AandB) The protein level of methylation-associated proteins (DNMT1, DNMT3a, DNMT3b, and Tet2) in aortas of ApoE−/− mice fed a Western diet (0, 6, 12, 18, and 24 weeks). Values are mean ± SEM from 6 animals in each group; *p < 0.05 vs. the 0-week group. (C and D) Protein level of methylation-associated proteins in vascular smooth muscle cells treated with TNF-α (5 or 10 ng/ml). Values are mean ± SEM of 3 samples in each group; *p < 0.05. (E and F) Protein-level alterations in DNMT1 and Tet2 induced by TNF-α in vascular smooth muscle cells were rescued with 5-aza treatment. Values are mean ± SEM of 3 samples in each group; *p < 0.05. (G) DNMT1 enzyme activity was measured in vascular smooth muscle cells treated with TNF-α or TNF-α + 5-aza. Values are mean ± SEM of 3 samples in each group; *p < 0.05. H, 5-hmC content, which represents Tet2 activity in vascular smooth muscle cells treated with TNF-α or TNF-α + 5-aza (n = 3). Values are mean ± SEM from 3 samples in each group; *p < 0.05. 5-hmC = 5-hydroxymethylcytosine; DNMT = deoxyribonucleic acid methyltransferase; Tet2 = ten-eleven translocation 2; other abbreviations as in Figure 2.

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