Fig. 2

Effects of 5-aza-2′-deoxycytidine (5-aza-dC) and trichostatin A (TSA) treatments on PLA2R1 promoter methylation (a) and PLA2R1 expression (b) in MDA-MB-453 cancer cells. a MS-HRM analysis of PLA2R1 methylations in control cells without treatment (one, green) and after exposure of MDA-MB-453 cells to 1 μM 5-aza-dC for 72 h (two, red), 0.3 μM TSA for 24 h (three, orange), and in cells treated with both agents (four, blue) as described in Materials and Methods section. Difference plots were normalized to standard 0 %-methylated DNA sample data. The black dotted lines represent standard xcurves of 0, 10, 25, 50, 75 and 100 % methylation degrees. Analyses were performed in duplicates and results are representative of two independent experiments. b Relative expression of PLA2R1-specific mRNA was determined using real-time RT-PCR and GAPDH-mRNA levels were used as reference in MDA-MB-453 cells. The experimental conditions were the same as described in (a). Values of comparative quantification were normalized to expression levels of untreated controls that were set at 1.0. Results are shown as means ± SD. Analyses were performed in triplicates and results are representative of two independent experiments