Figure 2From: Regulation of the stem cell marker CD133 is independent of promoter hypermethylation in human epithelial differentiation and cancerCD133 expression is repressed by DNA methylation in prostate cancer cell lines. (A) qRT-PCR analysis (relative to HPRT, calibrator = P4E6) of CD133 expression in prostate cell lines treated with 1 μM 5-Aza-2'-deoxycytidine for 96 hours. (N.D. = expression undetectable after 40 cycles, n = 2; ± SD, * = p < 0.05; ** = p < 0.01 in a paired t-test, p-value not calculated in cell lines with non-detectable levels of CD133). (B, C) FACs analysis of CD133 expression in prostate cell lines treated with 1 μM 5-Aza-2'-deoxycytidine for 96 hours. Live cells were stained with CD133/2(293C)-APC antibody (Miltenyi Biotec) (CD133-APC) or without any antibody (No Antibody) and analysed by FACs. For each dot plot, X axes: FITC channel fluorescence (not stained); Y axes: APC channel fluorescence (CD133 or No Antibody control); Percent of CD133+ cells is indicated and the results are summarised in C. (D) Dot-plot showing expression (qRT-PCR, relative to HPRT, calibrator = Bob) and methylation of CD133 in a panel of prostate cell lines grouped on the basis of their origin (benign or cancer) and their culture conditions (low FCS = 0%-2%; high FCS = 5%-10%).Back to article page