Fig. 4

Osteogenic differentiation in IL-32γ TG mice. a Calvarial cells isolated from wild-type (WT) or IL-32γ transgenic (TG) mice were cultured in osteogenic medium (OM). The cells were fixed and used for the measurement of alkaline phosphatase (ALP) activity or for staining with alizarin red (AR), or Von Kossa (VK) stains to assess the degree of OB differentiation and mineralization at 1, 2, 3, and 4 weeks (W). b WT or IL-32γ TG calvarial cells were stimulated with osteogenic media for 24 h and subjected to reverse transcription PCR analysis of the expression of regulatory genes needed for OB differentiation, including those that encode DKK-1, BMP-2, BMPRII, and LRP-5. c The DKK-1 protein level in the culture supernatant from the cells after 1 week of OB differentiation was determined by ELISA. Representative data from at least three independent experiments are shown; ***p < 0.001. BMP bone morphogenetic protein, BMPRII bone morphogenetic protein receptor II, DKK-1 Dickkopf-1, IL interleukin, LRP-5 low-density lipoprotein receptor-related protein 5