Methylation of PLA2R1 in DNA from the blood of a patient with AML after hematopoietic stem cell transplantation and minimal residual disease during three courses with azacitidine treatment. (A) Melt curve analyses of bisulfite-modified genomic DNA at the start (blue) and end (orange) of the first course of treatment, (B) at the start (blue) and end (orange) of the second course of treatment, and (C) at the start (blue) and end (orange) of the third course of treatment. (D) MS-HRM analyses at the start (blue) and end (orange) of the third course of azacitidine treatment. Melt curves of unmethylated (0%) and methylated (100%) standard DNA samples (A-C) and a standard curve with 0%, 5%, 10%, 20%, 30%, 50%, 75%, and 100% methylation ratios (D) are shown in black. (A-C); fluorescence dF/dT and (D) difference plot normalized minus 0. Analyses were performed in duplicates and results are representative of three independent measurements.