Epigenetic changes in the MMP2 gene promoter in MCF7 cells.

A: Sequencing of the MMP2 gene promoter. Closed and open circles represent methylated or unmethylated CpGs, respectively. On the left the number represent the sequenced clones. The 49 analyzed CpGs in the MMP2 promoter region are shown. Mock cells are at the top, above is 5-Aza-treated, hereafter FN-treated, recultured (MCF7 FN-treated and then cultured for 48 hours without fibronectin). The global methylation percentage is also shown at the right. B: Graphical analysis of CpG methylation pattern in the MMP2 promoter gene. The percentages of CpGs that were methylated in MCF7 mock (blue), MCF7 FN-treated (red) and MCF7 recultured (green) were shown. C: ChIP quantitative PCR analysis. Ct values were normalized between target and endogenous control (MMP2/GAPDH) and the results of mock (blue), FN-treated (red) and recultured (green) cells are shown. At the bottom the samples are separated according to the antibody used in the immunoprecipitation. Statistical comparison was performed using Student’s t Test. *p<0.05.