Figure 1.
Daple is required for coordinated ciliary beating in tracheal MCCs. (A) Schematic illustration of mucociliary clearance on the tracheal surface (upper panel) and immunofluorescence images of Odf2 (green) and Daple (magenta) in the tracheas of WT and Daple-KO mice (lower panels). (B) Analyses of mucociliary transport in isolated WT, Daple-KO, and Vangl1-KO adult mouse tracheas using live imaging of fluorescent beads. The bead position at each time point is presented as a color corresponding the time scale shown above the panel. Each bead was adjusted to a uniform starting time. Images are the magnified excerpts of boxes of micrographs shown in Fig. S1 F. (C) The ratios of end-to-end distances (d) against distances along the bead trajectory (D) of the fluorescent beads in arbitrarily selected regions of isolated tracheal surfaces from WT (n = 182), Daple-KO (n = 181), and Vangl1-KO (n = 71) mice. One-way ANOVA with Tukey-Kramer multiple-comparison test; **, P < 0.01. (D) Schematic illustration of cilia structures. The ciliary bases contain the BB marked as a ring (green) and the BF as a dot (magenta). The angle θi is between the horizontal axis and the vector originating from the center of the ith BB and ending at its BF. (E) Representative TEM images of WT and Daple-KO adult mouse tracheas. Yellow arrows indicate the direction of the BBs, and red arrows indicate abnormal orientation of BBs. High-magnification images of the boxed regions are shown in the lower panels. (F) Statistical analysis of the cell-level Io of the BB orientation in WT (n = 7 cells) and Daple-KO (n = 15 cells) adult tracheas. Each data point represents all BBs involved in one cell. Two-tailed Mann-Whitney U test; **, P < 0.01. (G) Representative immunofluorescence images of Odf2 and either centriolin or Cep128, respectively, with the BB marked as a ring (green) and the BF marked as a dot (magenta) in adult (n = 8 cells), P2 (n = 13 cells), and E17.5 (n = 10–12 cells) tracheas of WT and Daple-KO mice. High-magnification images of the boxed regions are shown with a line depicted from the center of the green ring to the center of the magenta dot and cell-level Io indicating the BB orientation in MCCs. The double arrows indicate the directions of the mounted tracheas in A, B, E, and G. Scale bars represent 5 µm in A, B, and G and 600 nm in E.

Daple is required for coordinated ciliary beating in tracheal MCCs. (A) Schematic illustration of mucociliary clearance on the tracheal surface (upper panel) and immunofluorescence images of Odf2 (green) and Daple (magenta) in the tracheas of WT and Daple-KO mice (lower panels). (B) Analyses of mucociliary transport in isolated WT, Daple-KO, and Vangl1-KO adult mouse tracheas using live imaging of fluorescent beads. The bead position at each time point is presented as a color corresponding the time scale shown above the panel. Each bead was adjusted to a uniform starting time. Images are the magnified excerpts of boxes of micrographs shown in Fig. S1 F. (C) The ratios of end-to-end distances (d) against distances along the bead trajectory (D) of the fluorescent beads in arbitrarily selected regions of isolated tracheal surfaces from WT (n = 182), Daple-KO (n = 181), and Vangl1-KO (n = 71) mice. One-way ANOVA with Tukey-Kramer multiple-comparison test; **, P < 0.01. (D) Schematic illustration of cilia structures. The ciliary bases contain the BB marked as a ring (green) and the BF as a dot (magenta). The angle θi is between the horizontal axis and the vector originating from the center of the ith BB and ending at its BF. (E) Representative TEM images of WT and Daple-KO adult mouse tracheas. Yellow arrows indicate the direction of the BBs, and red arrows indicate abnormal orientation of BBs. High-magnification images of the boxed regions are shown in the lower panels. (F) Statistical analysis of the cell-level Io of the BB orientation in WT (n = 7 cells) and Daple-KO (n = 15 cells) adult tracheas. Each data point represents all BBs involved in one cell. Two-tailed Mann-Whitney U test; **, P < 0.01. (G) Representative immunofluorescence images of Odf2 and either centriolin or Cep128, respectively, with the BB marked as a ring (green) and the BF marked as a dot (magenta) in adult (n = 8 cells), P2 (n = 13 cells), and E17.5 (n = 10–12 cells) tracheas of WT and Daple-KO mice. High-magnification images of the boxed regions are shown with a line depicted from the center of the green ring to the center of the magenta dot and cell-level Io indicating the BB orientation in MCCs. The double arrows indicate the directions of the mounted tracheas in A, B, E, and G. Scale bars represent 5 µm in A, B, and G and 600 nm in E.

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