Figure 6.

VCX2 expression in breast cancer cells is associated with loss of promoter methylation. The level of VCX2 promoter methylation in peripheral blood mononuclear cells (PBMCs) and breast cancer cells lines was investigated using bisulfite conversion followed by PCR melting point analysis. (A) The methylation level in PBMCs was similar to that of in vitro methylated DNA (IVM) and distinct from that of unmethylated DNA prepared by whole genome amplification (WGA). (B) The analysis was performed on cells with (T-47-D) or without (MDA-MB-231) endogenous expression of VCX2 and on cells with guadecitabine/valproic acid-induced expression of VCX2 [MDA-MB-231 + guadecitabine (Gua)/valproic acid (VPA)], which demonstrated inverse correlation between VCX2 promoter methylation and gene expression. Two different promoter regions were analyzed with highly similar results (only data for region 2 are shown). The experiment was performed with two biological replicates.