CCDC61 Associates with Basal Bodies and Plays a Role in Ciliogenesis
(A) xCCDC61 associates with basal bodies and rootlets in multi-ciliated epidermal cells of Xenopus embryos. A fluorescent image of a Xenopus embryo expressing xCCDC61-RFP (red), the basal body component Centrin2-BFP (blue), and the rootlet component Clamp-GFP (green). Scale bar, 3 μm.
(B) Location of hCCDC61 at the periphery of basal bodies of primary cilia. Immunofluorescent image of an RPE-1 cell transiently overexpressing GFP-hCCDC61. Co-immunofluorescent staining was performed against GFP (green), basal bodies (γ-tubulin, red), and the ciliary axoneme (ARL13B, magenta). Scale bar, 1 μm.
(C) Ciliated cells of control and CCDC61-knockout RPE-1 cells. Immunofluorescent images show representative immunofluorescent images used for quantifications of ciliogenesis of primary cilia. Scale bar, 10 μm. The bar graph shows that ciliogenesis was delayed in the CCDC61 knockout cells. Data shown correspond to three biological replicates (total cell counts n = 1,181, 1,103, and 1,008 for control, clone 1 and clone 2 cells after 24-h serum starvation respectively, and n = 1,151, 1,046 and 1,242 for control, clone 1 and clone 2 after 48-h serum starvation, respectively). Percentages are relative to control cells. Bar graphs show mean ± standard deviation.
See also Figure S5.