FIGURE 8.

Knockdown of Jmjd3 suppressed the transcriptional activity of Runx2 and osterix. A and B, schematic illustration of the promoter regions of Runx2 and osterix. The context of luciferase reporter vectors and the primers used for ChIP assay are indicated. C and D, the activity of Runx2 promoter, Runx2 3×binding site, and osterix promoter was decreased in shJmjd3 cells. E and F, knockdown of Jmjd3 increased the level of H3K27me3 on the Runx2 (E) and osterix (F) promoters. The shCont and shJmjd3 cells were cultured for 3 days in the osteoblast differentiation medium. Chromatin solution from the cells was subjected to ChIP analysis using anti-H3K27me3 and anti-IgG antibodies. PCR was performed with the primer pairs amplifying the promoter regions of Runx2 and osterix. IP, immunoprecipitate; C, shCont; J, shJmjd3.