Figure 7. Inhibition of Hh signaling attenuates Gli-1nLacZ reporter activity in glenoid fossa/eminence tissues and chondrocyte maturation in primary glenoid fossa cells.

Glenoid fossa/eminence from 2 month-old-Gli1-nLacZ reporter mice were culture in the presence of HhAntag at indicated concentrations (B, C) or control vehicle (A), processed for whole-mount LacZ staining (A–C) and subsequent sectioning (D, E). Note decreased LacZ activity in the articular cartilage of glenoid fossa/eminence treated with HhAntag vs control (D, E; respectively). Primary glenoid fossa cells were cultured on 24-well plates and treated with HhAntag at indicated concentrations or control vehicle for 9 days, fixed, and processed for Alkaline phosphatase activity (F, top panel) or Alizarin red staining (F, lower panel). Integrated density of Alkaline phosphatase-staining (G) and Alizarin red-staining (H) were quantified by ImageJ (G; n=3, *p <0.05, **p <0.02). Untreated cultures served as reference for comparison between groups. The data are expressed in arbitrary units. Histograms depicting HhAntag-dose dependent decrease of Hh signaling molecules and chondrocyte maturation markers in day 9 primary glenoid fossa cell culture compared to controls (**p <0.02). Scale bars: 2.4 mm in A for A–C; 80 μm in D for D–E. gf, glenoid fossa; zp, zygomatic process; ac, articular cartilage.