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. 2018 May 8;7(10):e008281. doi: 10.1161/JAHA.117.008281

Figure 4.

Figure 4

Effect of DNA methylation on the CYP11B2 promoter activity. A, Effect of complete CpG methylation. B, Effect of incomplete CpG methylation. C, Effect of angiotensin II. Bisulfite sequencing determined CpG methylation ratios. A methylation ratio at CpG5, dotted cycle was not able to be measured. Different luciferase constructs were methylated (filled circles) or mock‐methylated (open circles) in vitro and were transfected into H295R cells. Filled circles denote methylated cytosine residues; open circles, mock‐methylated. Results are given as luciferase activity normalized to cotransfected pRL‐TK Renilla luciferase activity. Results were mean±SEM of triplicate replicates. The numbering was based on the transcription start site of the CYP11B2 promoter. Complete CpG methylation totally abolished luciferase expression levels of methylated constructs (A). *P<0.01 vs incomplete methylation (B). *P<0.01 vs unmethylated DNA constructs (C). # P<0.01 vs compared 0.1 or 10 nmol/L of angiotensin II (Figure 2C).