Figure 8.
Rab11 reinforces the apical AJs during ventral furrow formation. (A) Capturing apical Rab11 vesicle dynamics with single-plane fast imaging. Example of a single constricting cell is shown. Scale bar, 5 μm. (A’) Zoom-in view of A showing two Rab11 vesicles moving toward an adherens junction spot. Scale bars, 1 μm. (B–D) Analysis of spatial correlation between Rab11 vesicles and AJs. (B) The same constricting cell as shown in A is outlined in green. Scale bars, 5 μm. Magenta circle: an example sampling region within which Rab11 signal is measured. Right panel: the intensity profiles of E-cadherin and Rab11 along the indicated cell outline. (C) E-cadherin intensities for all points along the cell outline plotted against the corresponding Rab11 intensities. (D) Rab11 shows a significantly higher correlation with E-cadherin than with a general membrane marker P4M (N = 3 embryos for each genotype, five cells per embryo. Cells from the same genotype were pooled together for statistical comparison). Error bars stand for SD. Two-sample, two-tailed unpaired t test was used for statistical comparison. **, P < 0.01. (E) Injection of Rab11DN rapidly eliminates apical Rab11 vesicles but does not immediately affect the perinuclear Rab11 compartments (green arrows). Scale bars, 10 μm. (E’) Zoom-in view of E. Scale bars, 2 μm. (F) Rab11DN injection affects the integrity of apical AJs in the constricting cells. 14 out of 19 Rab11DN injected embryos show fragmented apical AJs while eight of eight GST injected embryos show relatively continuous AJs. Scale bars, 5 μm. (G) Ajub shows reduced enrichment at the apical AJs during apical constriction in Rab11DN injected embryos compared to the control embryos (N = 4 embryos for each condition). Scale bars, 5 μm. Bottom, enlarged view of one cell (cyan boxes) as an example. Scale bars, 1 μm.

Rab11 reinforces the apical AJs during ventral furrow formation. (A) Capturing apical Rab11 vesicle dynamics with single-plane fast imaging. Example of a single constricting cell is shown. Scale bar, 5 μm. (A’) Zoom-in view of A showing two Rab11 vesicles moving toward an adherens junction spot. Scale bars, 1 μm. (B–D) Analysis of spatial correlation between Rab11 vesicles and AJs. (B) The same constricting cell as shown in A is outlined in green. Scale bars, 5 μm. Magenta circle: an example sampling region within which Rab11 signal is measured. Right panel: the intensity profiles of E-cadherin and Rab11 along the indicated cell outline. (C) E-cadherin intensities for all points along the cell outline plotted against the corresponding Rab11 intensities. (D) Rab11 shows a significantly higher correlation with E-cadherin than with a general membrane marker P4M (N = 3 embryos for each genotype, five cells per embryo. Cells from the same genotype were pooled together for statistical comparison). Error bars stand for SD. Two-sample, two-tailed unpaired t test was used for statistical comparison. **, P < 0.01. (E) Injection of Rab11DN rapidly eliminates apical Rab11 vesicles but does not immediately affect the perinuclear Rab11 compartments (green arrows). Scale bars, 10 μm. (E’) Zoom-in view of E. Scale bars, 2 μm. (F) Rab11DN injection affects the integrity of apical AJs in the constricting cells. 14 out of 19 Rab11DN injected embryos show fragmented apical AJs while eight of eight GST injected embryos show relatively continuous AJs. Scale bars, 5 μm. (G) Ajub shows reduced enrichment at the apical AJs during apical constriction in Rab11DN injected embryos compared to the control embryos (N = 4 embryos for each condition). Scale bars, 5 μm. Bottom, enlarged view of one cell (cyan boxes) as an example. Scale bars, 1 μm.

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