Fig. 2

Inhibited osteogenic differentiation potential of C3H10T1/2 cells with azoramide treatment in vitro. a, b Alkaline phosphatase (ALP) staining and quantification of ALP activity of cellular extracts subjected to different concentrations of azoramide (Azo). Scale bar = 100 μm. c, d Alizarin red S staining for mineral deposition and its quantification with different concentrations of azoramide. Scale bar = 100 μm. e Real-time qPCR of Runt-related transcription factor 2 (Runx2), Sp7, integrin binding sialoprotein (Ibsp), and bone gamma-carboxyglutamate protein (Bglap) mRNA in C3H10T1/2 cells subjected to 15 μM azoramide. f Western blot of Runx2 protein in C3H10T1/2 cells subjected to 15 μM azoramide. *p < 0.05, **p < 0.01, azoramide versus vehicle