Fig. 4

CBX4 suppressed Bmp2 activation of the Runx2 pathway by stabilizing the nuclear localization of HDAC3. A Small-ubiquitin-like modifier ligases CBX4, PIAS1-4, and RanBP2 in ACP cells treated with 200 ng/ml Bmp2 for 10 days were detected by Western blotting. B The flag-labeled CBX4 gene was transfected into ACP cells, and flag and CBX4 protein expression was quantified. ACP cells with and without flag-CBX4 over-expression were treated with Bmp2 (200 ng/ml) for 10 days, and nuclear localization of HDAC3 was detected by Western blotting (C), D immunofluorescence and E a dual-luciferase reporter system. F Western blotting was used to measure Runx2 protein levels. G–K The mRNA levels of Osterix, OCN, OPN, and ALP were determined by PCR. *P < 0.05