Figure 5.
Rab11 vesicles are transported apical–basally with a strong bias in the apical direction. (A) Imaging configuration to capture the movement of Rab11 vesicles along the apical–basal direction. Red line: imaging plane. Green outlines: constricting cells being imaged. (B and B’) Surface view of an embryo imaged as depicted in A. Scale bar, 10 μm. (B’) Zoom-in view of B showing the apical movement of two Rab11 vesicles (white circles). Scale bar, 2 μm. (C) Vesicle trajectories in three wildtype embryos in a ∼35-s time window. Red and blue mark apical- and basal-bound trajectories, respectively. (D) Counts of apically and basally oriented trajectories. (E) Comparison of average trajectory length, velocity, and duration (mean ± SD) shows no significant difference between apically and basally directed transport. Two-sample, two-tailed unpaired t test was used for statistical comparison; N = 3 embryos. (F) Left: Enface view of an embryo expressing both myosin and membrane markers. Right: Cross-section view of the same embryo at the indicated A-P positions (cyan lines). White line: imaging plane; yellow and red outlines: cells with or without obvious apical shrinking, respectively. Scale bars, 10 μm. (G) Rab11 vesicles exhibit directional bias toward the apical side. WT (cells with apical Myosin II activation): quantification of movies with Myosin II and Rab11 markers. The ROI may include a mixture of apically constricted and non-constricted cells that exhibited apical myosin accumulation. WT (constricting cell only): quantification of movies with membrane and Rab11 markers. Only apically constricted cells are quantified (Materials and methods). One-sample, two-tailed t test against 50% was used for comparison to hypothetical unbiased transport. Two-sample, two-tailed, unpaired t test was used for comparison between two WT samples. N = 3 embryos for each category. For all panels, error bar: SD; ***, P < 0.001; **, P < 0.01; *, P < 0.05; n.s., P > 0.05.

Rab11 vesicles are transported apical–basally with a strong bias in the apical direction. (A) Imaging configuration to capture the movement of Rab11 vesicles along the apical–basal direction. Red line: imaging plane. Green outlines: constricting cells being imaged. (B and B’) Surface view of an embryo imaged as depicted in A. Scale bar, 10 μm. (B’) Zoom-in view of B showing the apical movement of two Rab11 vesicles (white circles). Scale bar, 2 μm. (C) Vesicle trajectories in three wildtype embryos in a ∼35-s time window. Red and blue mark apical- and basal-bound trajectories, respectively. (D) Counts of apically and basally oriented trajectories. (E) Comparison of average trajectory length, velocity, and duration (mean ± SD) shows no significant difference between apically and basally directed transport. Two-sample, two-tailed unpaired t test was used for statistical comparison; N = 3 embryos. (F) Left: Enface view of an embryo expressing both myosin and membrane markers. Right: Cross-section view of the same embryo at the indicated A-P positions (cyan lines). White line: imaging plane; yellow and red outlines: cells with or without obvious apical shrinking, respectively. Scale bars, 10 μm. (G) Rab11 vesicles exhibit directional bias toward the apical side. WT (cells with apical Myosin II activation): quantification of movies with Myosin II and Rab11 markers. The ROI may include a mixture of apically constricted and non-constricted cells that exhibited apical myosin accumulation. WT (constricting cell only): quantification of movies with membrane and Rab11 markers. Only apically constricted cells are quantified (Materials and methods). One-sample, two-tailed t test against 50% was used for comparison to hypothetical unbiased transport. Two-sample, two-tailed, unpaired t test was used for comparison between two WT samples. N = 3 embryos for each category. For all panels, error bar: SD; ***, P < 0.001; **, P < 0.01; *, P < 0.05; n.s., P > 0.05.

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