Fig 8. Cilium-localized CK1γ promotes Smo phosphorylation and Shh pathway activity.

(A) NIH3T3 transfected with EYFP-CK1γ or EYFP-CK1γ-ΔC were immunostained with antibodies against acetylated tubulin (Ac-Tub), GFP, and Hoechst. EYFP-CK1γ but not EYFP-CK1γ-ΔC was localized on the primary cilium. (B–D) Gli-luciferase assay in NIH3T3 cells transfected Myc-Smo and the indicated CK1γ expression constructs in the presence or absence of DN-Kif3b, and treated with or without Shh. Data are means ± SD of normalized Gli-luc activity from three independent experiments. P-values are Student's t tests. (E–G) NIH3T3 cells were transfected Myc-Smo and the indicated CK1γ expression constructs in the presence or absence of DN-Kif3b, and treated with or without Shh. Cell lysates were immunoprecipitated with Myc antibody followed by western blot analysis with Myc and PS1 antibodies. (H, I) NIH3T3 cells were transfected with Myc-Smo and the indicated CK1γ expression constructs in the presence or absence of DN-Kif3b and treated with or without Shh. Cell lysates were immunoprecipitated with Myc antibody followed by western blot analysis with GFP and Myc antibodies. (J) NIH3T3 cells were transfected with the indicated constructs and treated with or without Shh or SAG, followed by immunoprecipitation and western blot analysis with the indicated antibodies.