Figure 2:Methylation of the PAI-1 gene in prostate cancer cell lines studied by MSRE-qPCR in comparison to MS-HRM analysis . (A) CpG sites and cut position of HhaI in the human PAI-1 gene. Sequence covered from position -50 to +200. The CpG sites are numbered relative to the transcription start site (TSS): CpG #1: 116; CpG #2: 109; CpG #3: 104; CpG #4: 91; CpG #5: 88; CpG #6: 49; CpG #7: 29. Horizontal bars represent the length of products obtained with MSRE and MS-HRM-qPCR. (B) Semi-quantitative results for PAI-1 gene methylation in normal and prostate cancer cells obtained by electrophoresis after MSRE-qPCR. (+) Digestion with HhaI, (-) control reaction without HhaI. (C) Methylation of the PAI-1 gene in normal prostate epithelial cells and prostate cancer cell lines analyzed with different methods. MS-HRM: analysis of CpG sites #1-7; MSRE: analysis of CpG site #7 (Figure 2A). Correlation index r was determined using SigmaPlot 11.2. Statistical difference was calculated by Mann-Whitney rank sum test. All analyses were performed in duplicates.