Effects of 5-aza-2´-deoxycytidine (5-aza-dC) treatment on PLA2R1 methylation (A) and PLA2R1 expression (B) in Jurkat and U937 leukemic cells. After exposure of cells to vehicle or 1 μM 5-aza-dC for 72 h mRNA and DNA were isolated. (A) MS-HRM analyses of PLA2R1 methylations with a standard curve of 0%, 25%, 50%, 75%, and 100% methylation (in black lines) without (in blue) and with exposure (in red) of cells to the DNA demethylating agent, 5-aza-dC. Difference plots normalized to the 0%-methylated standard DNA sample are shown. (B) Agarose gel electrophoresis of RT-PCR amplificates of PLA2R1 in comparison to TBP-specific mRNA in Jurkat and U937 cells, with and without exposure to 5-aza-dC. Lanes 1 and 2; control cells (without addition of 5-aza-dC), lanes 3 and 4 represent cells treated respectively with 5-aza-dC, L; ladder. Analyses were performed in duplicates and results are representative of three independent experiments.