Figure 5.
MT motor proteins regulate THN speed.(A) Both Brefeldin A and Dynasore cause a reduction in THN motility. Colored dots follow individual examples. Scale bars represent 20 µm in the overview and 10 µm in magnified images. Elapsed time in minutes. MHB, solid line; URL, dotted line. See also Video 5. (B and C) The overexpression of Kif1C-GFP leads to the accumulation of protein in the tips of phase 1 (B) and phase 2 (C) THNs. Scale bars represent 20 µm in the overview and 5 µm in magnified images. MHB, solid line; URL, dotted line. (D) Quantification of THN speeds confirms the loss of motility when endosomes or Golgi vesicle traffic is targeted. n = 20 embryos/164 tracks for drug control; n = 33 embryos/367 tracks for Brefeldin A; n = 23 embryos/162 tracks for Dynasore; n = 24 embryos/145 tracks for injection control; n = 24 embryos/170 tracks for Rab11a-expressing embryos; n = 25 embryos/252 tracks for Rab11a S25N-expressing embryos; n = 15 embryos/121 tracks for Kif1C-expressing embryos; n = 27 embryos/259 tracks for Kif1C T106N-expressing embryos; P = 9.56 × 10−16 control/Brefeldin A, P = 1.26 × 10−4 control/Dynasore, P = 0.001 control/Rab11a S25N, P = 9.6 × 10−3 Rab11a/Rab11a S25N, and P = 0.018 control/Kif1C T94N T106N. For full statistics, see Table S1. (E) Some Cdh2-positive particles appear to travel along MTs bidirectionally. Arrows indicate anterograde and arrowheads retrograde movement. Scale bar, 3 µm. Elapsed time in seconds on the right. (F) Cdh2 particles in THNs exhibit different motility patterns. Scale bar, 5 µm. Dotted line indicates region for kymograph depicted in G. The unidirectional EB3-GFP signals determine the classification of Cdh2 particles moving into antero- or retrograde direction, indicated above the images. Horizontal scale bars, 5 µm; vertical bars, 20 s. See also Video 6. (H) The kymographs illustrate that Cdh2 particle are less motile when either dynein activation is inhibited (p50) or anterograde vesicle traffic for Cdh2 is blocked (Kif5C T94N). Horizontal scale bars, 5 µm; vertical bars, 20 s. See also Video 6. Boxes in graphs represent 25–75% of all values and whiskers 1.5 times the quartile. Median is shown as a horizontal bar and mean as a square box. Significance level in Kruskal–Wallis ANOVA: *, P < 0.05; **, P < 0.01; ***, P < 0.001.

MT motor proteins regulate THN speed. (A) Both Brefeldin A and Dynasore cause a reduction in THN motility. Colored dots follow individual examples. Scale bars represent 20 µm in the overview and 10 µm in magnified images. Elapsed time in minutes. MHB, solid line; URL, dotted line. See also Video 5. (B and C) The overexpression of Kif1C-GFP leads to the accumulation of protein in the tips of phase 1 (B) and phase 2 (C) THNs. Scale bars represent 20 µm in the overview and 5 µm in magnified images. MHB, solid line; URL, dotted line. (D) Quantification of THN speeds confirms the loss of motility when endosomes or Golgi vesicle traffic is targeted. n = 20 embryos/164 tracks for drug control; n = 33 embryos/367 tracks for Brefeldin A; n = 23 embryos/162 tracks for Dynasore; n = 24 embryos/145 tracks for injection control; n = 24 embryos/170 tracks for Rab11a-expressing embryos; n = 25 embryos/252 tracks for Rab11a S25N-expressing embryos; n = 15 embryos/121 tracks for Kif1C-expressing embryos; n = 27 embryos/259 tracks for Kif1C T106N-expressing embryos; P = 9.56 × 10−16 control/Brefeldin A, P = 1.26 × 10−4 control/Dynasore, P = 0.001 control/Rab11a S25N, P = 9.6 × 10−3 Rab11a/Rab11a S25N, and P = 0.018 control/Kif1C T94N T106N. For full statistics, see Table S1. (E) Some Cdh2-positive particles appear to travel along MTs bidirectionally. Arrows indicate anterograde and arrowheads retrograde movement. Scale bar, 3 µm. Elapsed time in seconds on the right. (F) Cdh2 particles in THNs exhibit different motility patterns. Scale bar, 5 µm. Dotted line indicates region for kymograph depicted in G. The unidirectional EB3-GFP signals determine the classification of Cdh2 particles moving into antero- or retrograde direction, indicated above the images. Horizontal scale bars, 5 µm; vertical bars, 20 s. See also Video 6. (H) The kymographs illustrate that Cdh2 particle are less motile when either dynein activation is inhibited (p50) or anterograde vesicle traffic for Cdh2 is blocked (Kif5C T94N). Horizontal scale bars, 5 µm; vertical bars, 20 s. See also Video 6. Boxes in graphs represent 25–75% of all values and whiskers 1.5 times the quartile. Median is shown as a horizontal bar and mean as a square box. Significance level in Kruskal–Wallis ANOVA: *, P < 0.05; **, P < 0.01; ***, P < 0.001.

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