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. 2017 Oct 30;292(51):20859–20870. doi: 10.1074/jbc.M117.808022

Figure 6.

Figure 6.

Effect of DNA secondary structures in the CTCF-binding site on hTERT gene expression. a, schematic diagrams of the CpG free reporters containing WT promoter and mutants. b, transcriptional activities of reporters containing the WT or mutated hTERT promoters with hairpin structure (m1 and m3) or quadruplex structure (m2 and m4) were analyzed in A375 cells. The luciferase activities of reporters with mutated promoters were compared with that of WT. c, binding of CTCF to the reporters with WT or mutated promoters was further analyzed by ChIP assay using antibody against CTCF. The results represent the mean ± S.D. from three independent experiments. **, p < 0.01; ***, p < 0.001 compared with WT. d, EMSA experiments were performed to verify the binding of CTCF to biotinylated hT25, hT25-m1, hT25-m2, hT25-Me(1), hT25-Me(4), and hT25-m1-Me incubated with CTCF in vitro. Representative results from three independent experiments are shown. e, EMSA experiments were performed in the presence of KCl or LiCl condition for biotinylated hT25, hT25-Me(4), hT25-m1, and hT25-m2 incubated with CTCF in vitro.