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. 2015 Oct 7;5:14838. doi: 10.1038/srep14838

Figure 4. CTCF regulates transcriptional activation of the XAF1 gene.

Figure 4

(a) ACHN cells were pre-treated with 5-aza-2′-deoxycytidine (5 μM) and Trichostatin-A (0.2 μM) for 3 days. After that, the cells were transiently transfected with CTCF siRNAs or control scramble siRNAs. qPCR analyses were performed to measure the expression of both XAF1 and CTCF mRNA. HPRT was used as loading control. The means from three independent experiments were plotted with +SEM, *P < 0.05. (b) MCF-7 cells were transitorily co-transfected with both Wild-type-XAF1-promoter-SEAP or Δ-CTCF-XAF1-promoter-SEAP constructs and pMetLuc, which was used for transfection normalization. Data are represented as the means + SEM from three independent experiments, *P < 0.05. (c) MCF-7 stable clones of CTCF/Tet-On were stimulated with tetracycline at the indicated concentrations. Using qPCR assays, XAF1 and CTCF mRNA expression was normalized to HPRT, used as a loading control. The mean and range were plotted from two independent stable cell lines. (d) MCF-7-CTCF/Tet-On and MCF-7 Empty/Tet-On cell lines were transitory co-transfected with Wild-type-XAF1-promoter-SEAP and pMetLuc. After 48 h, tetracycline was added for 24 h. Data are represented as the means + SEM from three independent experiments, *P < 0.05. 5-Aza-2′-deoxycytidine (5-A-DC); Trichostatin-A (TSA); CTCF-binding site (CBS).