FIGURE 3.

Smo dependence of Shh signaling-induced localization of Sufu in primary cilia. A, representative immunofluorescent staining of Sufu (red) at the tips of primary cilia or B, quantification thereof in Smo^Flox/Flox MEFs infected with mock solution or Ad-cre viruses. Viral infection was for 12 h, and thereafter the cells were treated with either ShhN ligand or purmorphamine (Purm) for 24 h as indicated. “n” denotes the total number of primary cilia counted at each data point. Primary cilia were marked with anti-acetylated α-tubulin staining (green). C, representative immunofluorescent staining of Sufu at the tip of the primary cilium in Ptch^−/− cells. Cyclopamine (CPA) treatment was carried out after the cell culture reached confluence. D, quantification of Sufu-positive primary cilia; E, intensity of Sufu in primary cilia affected by cyclopamine treatment. F, effect of MG132 and cyclopamine on the intensity of Sufu in primary cilia in Ptch^−/− cells. n/s, not statistically significant (p > 0.1).