Fig. 1: Establishing distal ciliary domain in vertebrate MCC. | Nature Communications

Fig. 1: Establishing distal ciliary domain in vertebrate MCC.

From: A protein complex in the extreme distal tip of vertebrate motile cilia controls their organization, length, and function

Fig. 1: Establishing distal ciliary domain in vertebrate MCC.The alternative text for this image may have been generated using AI.

A Schematic representations of the role of MCCs in the human brain, respiratory tract and reproductive system. The tailbud stage of Xenopus embryo is covered by muco-ciliary epithelium, which resembles mammalian MCCs. B Image of Xenopus MCC labeled with Spef1-RFP (magenta), GFP-Ccdc78 (green) and membrane-BFP (blue). Scale bar represents 10 μm. C Immunofluorescence image of human trachea epithelial cell (HTEC) stained with anti-acetylated tubulin (magenta), anti-CCDC78 (green) and anti-CENTRIN (gray). The image of single cilium scraped from HTEC is shown on the right. Scale bar represents 5 μm. D, E Imaging of mouse airway MCCs (D, D’) and oviduct MCCs (E, E’) stained with anti-acetylated tubulin (magenta) and anti-CCDC78 (green). Scale bars represent 5 μm. F Confocal image of cilia labeled with GFP-Ccdc78 (green) and Spef1-RFP (magenta), magnified view of the cilium on the right with cartoon. (F’) Quantification of fluorescent intensity along the axoneme shown in right, the mean intensity was normalized by average intensity (n = 40 cilia, 10 cells, 4 embryos, 2 experiments). Data are presented as mean values +/- SD. We quantified its localization in Xenopus by measuring signal intensity in the distal-most four microns of individual cilia, which allowed us to ignore confounding signals as crowded cilia lay cross one another more proximally on MCCs. G Structural illumination microscopy (SIM) image of cilia labeled with GFP-Ccdc78 (green) and Spef1-RFP (magenta), magnified view of the cilium on the right with cartoon. (G’) Quantification of fluorescent intensity along the axoneme shown in right (see Methods)(n = 50 cilia, 10cells, 3 embryos, 2 experiments). Data are presented as mean values +/- SD. H TEM image of negatively stained Xenopus embryo MCC cilia with schematic model of same cilium. The tip is in green, central pair (CP) in pink and microtubule doublet (MTD) in gray. I Localization of mScarlet3-Armc9 (magenta, arrow) with GFP-Cfap52 (green) in a single MCC with a magnified single cilium from same cell shown at right. J Localization of mScarlet3-Ccdc78 (green) with GFP-Cep104 (magenta, arrowhead and arrow) in a single MCC with a magnified single cilium from same cell shown on right. K Localization of mScarlet3-Ccdc78 (green) with GFP-Eb3 (magenta, arrowhead and arrow) in a single MCC with a magnified single cilium from same cell shown at right.

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