Figure 1

Schematic representation of CpG island in seladin-1 promoter and methylation status in H295R and SW13 cell lines. (Panel A) The large CpG island includes the translation start codon (ATG), spanning from -868 bp to +918 bp. The arrows F1 and R1 indicate primers which did not evidence any methylation sensitive sites. F2 and R2 are primers used for MSP and Q-MSP analysis. (Panel B) Methylation specific PCR performed in H295R and SW13 cell lines. Primers for methylated (upper gel) and unmethylated DNA (lower gel) recognized in both cell lines corresponding methylated and unmethylated seladin-1 promoter sequences. After 6 days of treatment with 5-Aza (+5Aza) we observed a reduction of methylated form in both cell lines in comparison to controls (-5Aza). Conversely, the intensity of bands corresponding to unmethylated DNA increased in intensity, confirming the demethylation of corresponding sequences. (NTC = no template control, MW = molecular weight marker, M = cell line methylated DNA, UM = cell line unmethylated DNA).