Figure 9.
Planar polarized MTs are organized by Daple through MT bundling. (A) Representative images of the reconstituted UHVEMT with an extracted model of MTs (green) in WT and Daple-KO mouse tracheas (left; shown in Fig. 3 G), and magnification for the yellow boxed regions of either both models (middle) or only the extracted model of MTs (right). Arrows indicate apical MTs bundled at the Fzd side of the AJC. (B) Stepwise bleaching of spots of mEGFP–Daple FL, CT, NT, and ΔCT molecules. (C) Cumulative histogram of initial fluorescence intensities of immobilized spots of mEGFP–Daple FL (n = 158), mEGFP–Daple CT (n = 196), mEGFP–Daple ΔCT (n = 213), and mEGFP–Daple NT (n = 129). (D) Schematic illustration of the MT bundling assay. The mixture of Daple and MTs was added to the flow chamber. Samples were imaged using TIRF microscopy. (E) Representative TIRF images of the MT bundling assay with mEGFP–Daple FL (or CT; green) and fluorescently labeled MTs (magenta). Yellow arrows indicate MTs to which Daple bound, and white arrows indicate MTs to which Daple did not bind. High-magnification images of the boxed regions are shown. (F) Representative images of negative staining EM of MTs alone (left panel), MTs with 10 µM Daple FL (middle panel), or MTs with 10 µM Daple CT (right panel). (G) Schematic drawing of the interaction between Daple and MTs at the Fzd side of the AJC. Daple directly binds to, bundles, and stabilizes MTs through its dimerization and accumulates apical MTs at the Fzd side of AJC in MCCs. Scale bars represent 250 nm in A, 1 µm in E, and 200 nm in F. A.U., arbitrary units.

Planar polarized MTs are organized by Daple through MT bundling. (A) Representative images of the reconstituted UHVEMT with an extracted model of MTs (green) in WT and Daple-KO mouse tracheas (left; shown in Fig. 3 G), and magnification for the yellow boxed regions of either both models (middle) or only the extracted model of MTs (right). Arrows indicate apical MTs bundled at the Fzd side of the AJC. (B) Stepwise bleaching of spots of mEGFP–Daple FL, CT, NT, and ΔCT molecules. (C) Cumulative histogram of initial fluorescence intensities of immobilized spots of mEGFP–Daple FL (n = 158), mEGFP–Daple CT (n = 196), mEGFP–Daple ΔCT (n = 213), and mEGFP–Daple NT (n = 129). (D) Schematic illustration of the MT bundling assay. The mixture of Daple and MTs was added to the flow chamber. Samples were imaged using TIRF microscopy. (E) Representative TIRF images of the MT bundling assay with mEGFP–Daple FL (or CT; green) and fluorescently labeled MTs (magenta). Yellow arrows indicate MTs to which Daple bound, and white arrows indicate MTs to which Daple did not bind. High-magnification images of the boxed regions are shown. (F) Representative images of negative staining EM of MTs alone (left panel), MTs with 10 µM Daple FL (middle panel), or MTs with 10 µM Daple CT (right panel). (G) Schematic drawing of the interaction between Daple and MTs at the Fzd side of the AJC. Daple directly binds to, bundles, and stabilizes MTs through its dimerization and accumulates apical MTs at the Fzd side of AJC in MCCs. Scale bars represent 250 nm in A, 1 µm in E, and 200 nm in F. A.U., arbitrary units.

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