Figure 2.

Cavarial cells from PHF2-t/g mice differentiated toward osteoblasts earlier than those from their wild-type littermates. Calvarial cells from wild-type and PHF2-t/g mice were cultured in an osteogenic medium containing BMP2 for the indicated times. (A) PHF2 and Runx2 levels were determined by immunoblotting. (B) Immunfluorescence analyses for PHF2 and Runx2 localization were performed in wild-type calvarial cells on differentiation day 1. (C, D) Osteoblast differentiation was monitored by staining ALP (C) and by measuring its activity (D). (E) Calvarial cells undergoing differentiation were stained with Alizarin to monitor mineralization. (F) The mRNA expressions levels of osteogenic genes (ALP, OCN, and BSP) were analyzed by RT-qPCR and were normalized to 18S RNA levels. The results (mean ± SD, n = 3) were presented as the relative values to those on day 0, and ^* denotes P < 0.05 versus the wild-type value on the corresponding day.