Figure 1

Epigenetic regulation of Trip10. (A) Bisulfite sequencing (left) and qMSP (right) shows TripP10 methylation in various cancer cell lines. CpG locations are indicated as vertical bars in the promoter and first exon of Trip10 (top). Arrows mark the location of MSP primers. Open circles indicate unmethylated CpG sites, and circles filled to varying degrees reveal the percentage of methylation at specific CpG sites. Results of eight clones from each cell line are presented. For qMSP, Col2A1 was used as loading control. (B) H3K4me3 and H3K27me3 association at Trip10 promoter were demonstrated by ChIP analysis. CREB, AML-1α, and ER transcription factor binding sites are shown with individual CpG sites (short vertical bars). Arrows indicate the bisulfite sequencing region shown in (A). All three transcription factor binding sites were associated with H3K4me3, but not H3K27me3. (C) DNA demethylation. IMR-32 cells treated with 5-Aza (20 μM) or DMSO (vehicle) were analyzed by qMSP and qRT-PCR. Col2A1 served as loading control for qMSP, and GAPDH served as loading control for qRT-PCR.