A. Strategy used to perform restriction enzyme accessibility assay of the uPA promoter. Genomic DNA of each cell line was digested with restriction enzyme PvuII before PCR amplification with a set of three primers (uPA-F1, uPA-F2, and uPA-R). The ratio of 925:241 bp products reflects promoter accessibility.
B. Left, RE accessibility of the uPA promoter region in human cancer cell lines SK-N-BE, SK-N-AS, SF-3061, LNCaP and PC3. Right, densitometry of undigested (− PvuII) and digested (+PvuII) PCR products. The decrease of the ratio 925:241 bp after PvuII digestion indicates that the chromatin is accessible to PvuII ( p <0.05).
C. Top, chromatin accessibility changes of the uPA promoter region in SK-N-BE, SK-N-AS, SF-3061 and LNCaP cells following treatment with TSA. Bottom, densitometric analysis of undigested (− PvuII) and digested (+PvuII) PCR products in control and TSA-treated cell lines. The ratio was decreased by the treatment of TSA (p <0.05).
(Results are representative of three separate experiments.)