Figure 2

WIF1 methylation in primary breast cancer. WIF1 methylation analyses of primary breast cancer specimens. MSP was performed on bisulfite-treated DNA from breast cancer (T) and matching normal primary breast tissues (N). MSP results from three representative matched pairs and eight additional breast carcinomas (#) are shown. DNA bands in lanes labeled with U indicate MSP products amplified with primers recognizing the unmethylated promoter sequence. DNA bands in lanes labeled with M represent amplified MSP products with methylation-specific primers. Peripheral blood lymphocytes (PBL) and breast cancer cell line ZR75-1 served as positive controls for the methylation-specific reaction, respectively. Water was used as template in the 'no template control' (NTC). Note that tumor tissue usually displayed a PCR product in the U-reaction as well, due to contaminating normal tissue (stromal cells, endothelial cells) present in the tumor specimens as has also been described by Suzuki et al. [34].