Figure 1.

Promotion of osteoblast differentiation by Ihh. (A) C3H10T1/2 cells were cultured with or without Indian hedgehog (Ihh; 500 ng/ml) or Sonic hedgehog (Shh; 500 ng/ml) for 7 d. The cells were determined by ALP activity and staining. Data represent mean ± SD. *p < 0.01. Similar results were obtained from five independent experiments. (B) C3H10T1/2 cells were infected with control, Gli2 or Gli3 adenovirus as indicated, and cultured for 7 d in the absence or presence of Ihh (500 ng/ml). The total RNA isolated from the cells was subjected to RT-PCR analysis using the specific primes for osteocalcin (top) or β-actin (bottom). Similar results were obtained from three independent experiments. (C) Primary mouse osteoblasts infected with control adenovirus were cultured in the presence or absence of either BMP2 (300 ng/ml) or Ihh (500 ng/ml). Primary mouse osteoblasts infected with Myc-tagged wild-type or dominant-negative Gli2 adenovirus (DN-Gli2) were incubated without BMP2 and Ihh. After a 14-d incubation, the cells were examined by Alizarin red staining. Similar results were obtained from three independent experiments.