Figure 5. Involvement of cilia-mediated Shh signaling in neuronal differentiation of OCRL-deficient iN cells.

(a, b and c) Quantitative real-time PCR (RT-PCR) validation of Shh signaling genes SHH, GLI1 and PTCH1 in iN cells. RT-PCR was repeated three times with different batches. Gene expression values are normalized to GAPDH. (d) Brain section of wild-type and IOB mouse stained with SHH (red) and Arl13b (green) antibodies. DNA stained with DAPI (blue). Scale bars as indicated. (e) Quantification of the percentage of positive ciliated cells. > 100 cells analyzed for each independent experiment. (f, g, h and i) Quantitative real-time PCR (RT-PCR) validation of Shh signaling genes Gli1, Gli2, Gli3 and Ptch1 in brain sections of wild-type and IOB mouse. RT-PCR was repeated three times with different batches. Gene expression values are normalized to actin. (j) Western blot analysis using antibodies against SHH, GLI1 and β-actin in brain sections of wild-type and IOB mouse. The bars in each graph represent mean ± SD. Statistical significance was determined using Student’s t-test, with exact p-values reported.