To confirm our MSP results and determine how ISL affects WIF1 promoter methylation in breast cancer, we used bisulfite sequencing to analyze the methylation status of 39 CpG sites within the 415-bp fragment of the WIF1 promoter (from −555 to −140) in MDA-MB-231 and MCF-7 cells (Figure 8B). This evidence concerns the gene WIF1 and breast carcinoma.