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. 2013 Jul 25;22(24):5036–5047. doi: 10.1093/hmg/ddt354

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Published by Oxford University Press 2013. This work is written by (a) US Government employee(s) and is in the public domain in the US.

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Figure 4. — Enhancer effect of the methylated APOE CGI in pCpG-free vector reporter assays. (A) The diagram of reporter constructs with an APOE CGI fragment (1483 bp) in either the forward or reverse orientation, 5′ of the hEF-1 promoter. (B) The APOE CGI was either methylated by DNA methyltransferase treatment (methylation section) or left un-methylated and transiently transfected into three cell lines (HepG2, SH-SY5Y and U118). AP activity of each construct was measured and compared with its promoter-only construct counterpart (set at 1.0). Each reporter activity was generated from five experimental replicates. Expression levels between constructs were compared by one-way ANOVA [Forward: HepG2, F(5, 78) = 0.54, P = 0.74; SH-SY5Y, F(5, 66) = 7.39, P < 0.001; U118, F(5, 48) = 4.04, P = 0.004; Reverse: HepG2, F(5, 84) = 0.98, P = 0.43; SH-SY5Y, F(5, 66) = 2.84, P = 0.02; U118, F(5, 48) = 4.56, P = 0.002]. Pair-wise comparisons were made with a post hoc Bonferroni test (*P < 0.05, **P < 0.01, ***P < 0.001).