FIGURE 6.

Recruitment of chromatin modifier SUZ12 and JMJD3 to EpCAM promoter in undifferentiated and differentiated hESCs. Top, schematic representation of the EpCAM promoter locus, which spanned positions −630 to +967 with respect to the TSS. The ChIP primers used in the study are indicated by horizontal lines. A and B, chromatin samples were immunoprecipitated with anti-SUZ12 antibody (A) or anti-JMJD3 antibody (B), and enrichment of the EpCAM and KRT1 promoter was quantitated by Q-PCR. KRT1 was used as a control for SUZ12/JMJD3/H3K27me3 binding. Each experiment was done in triplicate (mean ± S.D.). The value of immunoprecipitated target was calculated as the ratio of IP DNA to the total input DNA (IP/input). The target IP/input was further normalized to a control promoter region of HBB (SUZ12) or of GAPDH (JMJD3) to obtain -fold enrichment values. By ChIP measurement, the association of SUZ12 with the EpCAM promoter was elevated both upstream and downstream of TSS in differentiated H9 cells. In contrast, quantification of the intensities of JMJD3 binding was increased downstream of TSS in undifferentiated H9 cells (*, p < 0.05).