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. 2018 Dec 31;116(3):874–879. doi: 10.1073/pnas.1804042116

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Fig. 6. — Inversin determines Smo ciliary translocation for Shh pathway activation. (A and B) Inversin is required for the ciliary translocation of Smo. Ptc1^−/− or Ptc1^+/− cells were knocked down for Inversin (Invs^KD) and arrested at the G0 phase. After the indicated treatment, the cells were immunostained for the indicated proteins. Arrowheads indicate the ciliary base. The percentage of PC- and EC-localized Smo is shown. (C) Inversin is required for Shh signaling transduction. Ptc1^+/−, Ptc1^−/−, or Ptc1^+/− cells knocked down for Inversin were arrested at the G0 phase; after the indicated treatment, the cells were lysed and the indicated proteins examined. (D) Smo^M2 cannot translocate into the cilia of cells with Inversin knockdown. WT cells stably expressing Smo^M2-GFP were knocked down for Inversin, arrested at the G0 phase, and immunostained for the indicated proteins. The percentage of EC-localized Smo is shown. In the statistics results from A, B, and D, the values are the mean ± SD; 50 cells per sample were randomly selected and counted in each of three independent experiments. (E) Smo^M2 strongly interacts with Inversin in the absence of ShhN. HEK 293T cells were cotransfected with the indicated plasmids, arrested at the G0 phase, and processed to immunoprecipitation assays.