Fig. 4: Alteration of ciliary composition and cilia-related signaling in patient fibroblasts.

(a,b) Representative immunofluorescence images of fibroblasts stained for CEP164 (red) and ARL13B (green) in (a); INPP5E (red) and ARL13B (green) in (b). The arrowheads indicate cilia. Scale bars, 20 μm. (c,d) Quantification of relative levels of ARL13B (a and Fig. S7a) and INPP5E (b and Fig. S7b) in cilia. Statistical significance was determined by the t-test (**p < 0.01). (e,f) Quantitative real-time polymerase chain reaction (PCR) analysis of GLI1 (e) and PTCH1 (f) expression in fibroblasts. The cells were subjected to serum starvation for 48 hours followed by SAG treatment or no SAG treatment for 24 hours. Three biological repeats were used for each group. Statistical significance was determined by the t-test (**p < 0.01; ***p < 0.001; ns not significant). (g–i) Representative immunofluorescence images of fibroblasts stained for SMO (green) and ARL13B (red) in (g); GPR161 (red) and ARL13B (green) in (h,i). The cells were subjected to serum starvation for 48 hours followed by SAG treatment or no treatment for 24 hours. Scale bars are 20 μm in (g,h) and 3 μm in (i). (j–l) Quantification of cilia positive for SMO (g) and GPR161 (h, i). Statistical significance was determined by the t-test (***p < 0.001). Error bars represent SD.