Fig. 1. Rapid localization of Smo in primary cilia after activation of the Hh pathway and regulation by Ptc1. (A) Immunoblots with antibodies to Ptc1, Smo, and actin were used to assess amounts of endogenous proteins in extracts from NIH 3T3 cells treated with Shh or SAG (100 nM). (B) Enrichment of Smo in primary cilia of NIH 3T3 cells left untreated (control) or treated with Shh or SAG (100 nM) for 24 hours. (C) Mean intensity of Smo fluorescence in cilia of NIH 3T3 cells treated with Shh or SAG (100 nM). Each point shows the mean ± SEM of fluorescence from 10 to 20 cilia. (D and E) Constitutive presence of Smo in the cilia of unstimulated ptc1^–/– MEFs and reversal by retrovirally transduced Ptc1-YFP. In (B) and (D), confocal images of the ciliary marker acetylated tubulin (red) and Smo (green) were detected by immunofluorescence; nuclei (blue) were stained with 4',6'-diamidino-2-phenylindole (DAPI). [View Larger Version of this Image (234K JPEG file)]

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