Fig. 3

Bmp2 suppressed HDAC3 nuclear translocation to activate the Runx2 signaling pathway and promote ACP osteoblastic differentiation. A After HDAC3 protein levels were knocked down by siRNA, 200 ng/ml Bmp2 was used to treat ACP cells for 10 days, and the calcification was assessed by Alizarin red staining (B). C RGFP966 (10 µM), an inhibitor of HDAC3 activation, was used as pretreatment for 1 h and then Bmp2 (200 ng/ml) was added to cells with HDAC3 knocked down or expressed. Runx2 protein levels were analyzed by Western blotting. D–G The mRNA levels of Osterix, OCN, OPN, and ALP were examined by PCR. ACP cells were treated with 200 ng/ml Bmp2 for 10 days, and the nuclear translocation of HDAC3 was detected by Western blotting (H), immunofluorescence (I) and (J) a dual-luciferase reporter system. *P < 0.05