Figure 8.

KIF14 regulates HH signaling independently of AURA activity. (A–C) Functional analyses of HH pathway activation following control or KIF14 silencing and mock or 2 µM SAG treatment in nHDFs. (A) Quantitative RT-PCR quantification of the effect of GLI1 on mRNA levels. (B and C) Analysis of the effect of KIF14 depletion on SMO translocation to the ciliary axoneme and its intensity. (B) Representative images of SMO (green) and ARL13B (red) IF detection; scale bar, 2 µm. (C) Quantification of changes in relative SMO intensity. (D–F) Examination of effect of TCS7010 AURA inhibition on response of nHDF cells transfected with the indicated siRNA to HH pathway activation. Experimental setup was the same as in Fig. 7 A, but with an additional 0.5 µm SAG treatment for the last 24 h. (D) Representative images of SMO (green) and ARL13B (red) IF detection in nHDF cells transfected with ctrl or KIF14 siRNA and treated with mock or AURA inhibitor; scale bar = 2 µm. Quantification of changes in SMO intensity (normalized to ARL13B) is shown in E and quantitative RT-PCR quantification of effect on mRNA level of GLI1 in F. Asterisks or “ns” indicate statistical significance determined using Tukey's multiple comparisons test.