FIGURE 3.
Inhibition of hedgehog signaling with cyclopamine rescues the chondrogenic defect in Prx1-Cre:Ptc1c/c micromass. For micromass experiments, cells were harvested from 11.5 dpc autopods and maintained in culture for up to 8 days. Micromass cultures derived from Prx1-Cre:Ptc1c/c limbs (I–P) lacked the Alcian blue staining profile observed in control cultures (A–H). Addition of cyclopamine disrupted nodule morphology in control cultures (A′–H′) and restored Alcian blue staining in Prx1-Cre:Ptc1c/c micromass (I′–P′). ImageJ analysis shows the Alcian blue-stained area as a percentage of the total micromass area. Results are representative of three individual micromass experiments. Scale bar, 1 mm. Addition of cyclopamine (5 μm) significantly reduced both Gli1 and Ptc1 expression levels in day 2 and 4 micromass cultures from Prx1-Cre:Ptc1c/c limbs (Q–T) (*, p < 0.05). Error bars represent S.D.