Figure 6. CC develops in Cep290 mutant prior to degeneration.

(A–H) SIM images (high magnification, right; low magnification, left) of cilia from P10 WT, rd16, NN, and KO animals labeled with CEP290, AcTub, and centrin. Scale bar on left: 1 μm. SIM (right) images of representative cilia. Row average intensity and column average intensity plots are shown. AcTub and centrin gains were adjusted to subsaturation for image presentation. CEP290 intensity levels in Cep290 mutants were normalized to WT levels. (I and J) Dot plots with averages and standard deviations of the maximum radii and lengths of CEP290, AcTub, and centrin in the cilium. Cilia were imaged from 3 nonlittermate mice for each genotype. Thirty-three percent of the maximum intensity value of each channel was used as the boundary criterion for the measurement of each cilium. Measurements were compared with 1-way ANOVA and Dunnett’s post hoc test. (K) Western blots demonstrating detection of CEP290 protein products. Expected sizes for WT CEP290, CEP290 (rd16), and AcTub are 290 kDa, 270 kDa, and 50 kDa, respectively. The N-terminal blot lanes were run simultaneously on the same samples as the C-terminal blot. Asterisk indicates band shift in the rd16 protein product. Dotted line indicates CC/OS border. Full blot for K is available in the supplemental materials. AcTub, acetylated α-tubulin; CC, connecting cilium; WT, wild-type; rd16, Cep290^rd16; nn, near null-Cep290^tm1.1Jgg; ko, Cep290-knockout; ND, not detectable.