Fig. 5

Azacytidine induces demethylation and expression of BRF2 selectively in MCF-7 cells. a, b, c MCF-7 and (d, e) MDA-MB-231 cells were treated with 5 μM 5-Azacytidine for 24, 48 and 72 h. Post-treatments, total RNA was isolated and analyzed by qRT-PCRfor expression of (a, d) BRF2 and (c, e) BRF1 mRNAs. The ΔΔCt method with GAPDH and RPS13 expression levels was used as a reference for normalization. Meta-analysis of two independent experiments performed in triplicate was completed using one-way ANOVA with a Tukey’s post-test with a 95 % confidence interval (Graphpad Prism 3.03); * = p <0.05; ** = p < 0.01; *** = p < 0.001. (b) Genomic methylation of BRF2 promoter DNA in MCF-7 cells was analyzed with methylation-sensitive restriction enzymes, as in Fig. 4a