FIGURE 3.

Increased transcript expression of the Pcdhβ gene cluster is not primarily mediated by altered promoter methylation. A, her-2 tumor cells were treated with the demethylating reagent 5-aza-2′-deoxycytidine at two different concentrations for 3 days, and total RNA was isolated and used for detection of transcript levels of Pcdhβ. For each transcript, the values are normalized to control (Gapdh or Rpl4) and expressed as means with error bars indicating ±1 S.D. of three independent experiments. B, after the genomic DNA samples were purified and bisulfate-treated, methylation-specific PCR for the Pcdhβ genes was performed in two wild-type (samples 1 and 2) and two GnT-V knock-out (samples 3 and 4) tumors. The presence of a PCR product band in lane M or U indicates methylated or unmethylated genes, respectively. Data are representative of two independent experiments.