Figure 6. Ift88^c.mut have fewer, shorter cilia throughout the cerebellum and mislocalization of ciliary membrane markers.

(A) Representative images illustrating cilia loss in the cerebellum of Ift88^c.mut animals, immunostained for ARL13B to label cilia (red), Pericentrin (PCNT) to label centrosomes (green) and nuclei (blue) Scale bar = 20 μm. (B) Western blot analysis of IFT88 in cerebellum lysate 3 months after TMX injections in Ift88^c.mut animals. (C) Quantification of cilia loss. Compared to Ttbk2^c.mut mice, cilia loss is less dramatic in the cerebellum of Ift88^c.mut animals (each point represents a field scored, 45 fields scored per genotype. n = 3 animals. p<0.0001 by student's unpaired t-test, error bars indicate SEM). (D) Quantification of cilia length between Control and Ift88^c.mut. Cilia in Ift88^c.mut cerebellum are shorter (each point represents a single cilium, 80 cilia were measured for each genotype. n = 3 animals, p<0.0001 by student’s unpaired t-test, error bars indicate SEM). (E,F) Cilia from 6-month-old Control and Ift88^c.mut stained for γ-Tubulin to label centrosomes (magenta), ARL13B to label cilia membrane (red), AC3 to label cilia membrane (green) and DAPI (blue). Ift88^c.mut lose AC3+ cilia. (E) The arrow indicates a cilium that is AC3+/ARL13B-. Scale bar = 5 μm (E) and 1 μm (F). (G) Quantification of AC3+ cilia throughout the cerebellum. Ift88^c.mut animals have a strong reduction in AC3+ cilia localization (each point represents a field scored, 36 field scored per genotype. n = 3 animals. p<0.0001 by student’s unpaired t-test, error bars indicate SEM).