Figure 1

UCHL1 promoter in RCC cell lines. A) Schematic diagram of the UCHL1 core promoter DNA region with its respective CpG islet. The sequence segment of interest taken from the reference GI 16949651 as indicated is displayed below the scheme. The putative methylation sites (CpG dinucleotides) are underlined in the sequence stretch. (B) Representative COBRA pattern for RCC cell lines displaying a distinct methylation status of the UCHL1 promoter DNA (MZ1851RC: unmethylated; MZ1851LN: fully methylated; MZ2862RC: partially methylated) are shown. Genomic DNA extracted from the given RCC cell lines upon treatment with different DAC concentrations was treated with bisulfite and amplified by nested PCR as described in Methods. The resulting 265 bp amplicons were either digested with BstU I (+) or left untreated (-) and subsequently separated in 2% agarose gels in TAE buffer. A 100 base pair DNA ruler loaded in the first lane served as length standard.