FIGURE 5.

Establishment of stable Jmjd3 knockdown cells. MC3T3-E1 cells were transfected with nonspecific (shCont) or Jmjd3-specific (shJmjd3) shRNA and cultured for 3 days in the osteoblast differentiation medium. A, the expression levels of Jmjd3 mRNA were examined by real-time PCR. Data are presented as means ± S.E. **, p < 0.01. B, Western blot analysis was performed using the specific antibodies for Jmjd3, β-actin, H3K27me3, and H3. The relative expressions of Jmjd3/β-actin and H3K27me3/H3 were calculated by computer software (ImageJ).