Figure 2

PHI induces p16 hypomethylation in RPMI8226 myeloma cells. The myeloma cells were cultured with or without phenylhexyl isothiocyanate (PHI) at varying concentrations for 10 days. Cellular DNA was extracted and bisulfite-converted as described in Material and Methods. Methylation-specific PCR was performed using primers specific for methylated and unmethylated DNA forms of p16, respectively. The PCR product was visualized after agarose gel electrophoresis. Decitabine (5-aza) was used as a positive control for DNA hypomethylation. M, methylated p16 fragment; U, unmethylated p16 fragment. The 150 bp marker position was indicated.