Figure S1.
Quantitation of cell and nuclear behaviors in control and myosin disrupted epithelia. (A and A′) Shape factor at the nuclear midplane for cell (blue) and nucleus (red) from 10 min prior to 20 min mid-GBE in wildtype embryos and a box chart showing the comparisons at −5 and 20 min of GBE in A′. (B) Ratio of major to minor axis length for cell (blue) and nucleus (red, same as in Fig. 2 B) at nuclear midplane from 10 min prior to 20 min mid-GBE in wildtype embryos. (B′) Major:minor axis ration at the nuclear midplane for cell (blue) and nucleus (red) at 0 and 20 min mid-GBE in wildtype embryos. For A–B′, n = 605 cells and nuclei from k = 5 embryos. For A and B, light blue shading indicates GBE. Error envelopes indicate SD. (C) Still images showing cell deformation is required for nuclear deformation and dispersion indicated by fairly round nuclei present in Y-27632–injected (Y27) embryos compared to vehicle-injected (control) embryos. (D) Comparison of the major:minor axis percent change at nuclear midplane for cell (blue) and nucleus (red) between control and Y27 embryos. The negative percent change indicates a decrease in the axis ratio and the positive percent change indicates an increase in the axis ratio; n = 186 cell and nuclei for control and n = 236 cell and nuclei for Y27 from k = 3 embryos for each background. (E) Example of T1 transition with a color overlay of T1 interface (blue) shared by cells A and B, T2 vertex (yellow) shared by cells A–D, and T3 interface (red) shared by cells C and D (top). Bottom panels show the centroid (white) distance between AB (blue) and CD (red) cells. (F) Absolute nuclear midplane positions in AB, CD, and All cells. (G) Peak velocity with which nuclei move in apical–basal direction measured pre- (−5 to 0 min) and mid- (+15 to +20 min) GBE in wildtype embryos; n = 213 nuclei for pre- and n = 512 for mid-GBE from k = 3 embryos. (H) Maximum apical–basal speed of nuclei in AB, CD, and All cells. For F and H, n = 36 for AB and CD cells and n = 82 for All cells from k = 1 embryo. (I and I′) Comparison of probability change in relative nuclear position at pre- (−5 to 0 min, blue), early (0–10 min, yellow) and mid- (15–20 min, red) GBE in control and Y27 embryos; n = 3,339 nucleus pairs for control and n = 5,565 nucleus pairs for Y27 embryos from k = 3 embryos for each background. (J) Heatmap showing apical–basal distribution of nuclei (absolute midplane position) over the course of GBE in Y27 embryos. Color bar indicates the probability of nuclear midplane position; n = 1,097 nuclei from k = 3 embryos. (K) Maximum packing ratio achieved by clusters of nuclei as measured by the dispersion metric at early (0 min) and mid- (+20 min) GBE; for control, n = 370 clusters of cells for early and 336 clusters of cells for the mid-GBE, for Y27, n = 803 clusters of cells from early and 870 clusters of cells from mid-GBE from k = 3 embryos for each background. The measured n values are regardless of time points where not indicated. For C and E, scale bar = 10 and 5 μm, respectively. Statistical significance was calculated using the Mann–Whitney U-test. ns = not significant, ***P < 0.001, ****P < 0.0001.

Quantitation of cell and nuclear behaviors in control and myosin disrupted epithelia. (A and A′) Shape factor at the nuclear midplane for cell (blue) and nucleus (red) from 10 min prior to 20 min mid-GBE in wildtype embryos and a box chart showing the comparisons at −5 and 20 min of GBE in A′. (B) Ratio of major to minor axis length for cell (blue) and nucleus (red, same as in Fig. 2 B) at nuclear midplane from 10 min prior to 20 min mid-GBE in wildtype embryos. (B′) Major:minor axis ration at the nuclear midplane for cell (blue) and nucleus (red) at 0 and 20 min mid-GBE in wildtype embryos. For A–B′, n = 605 cells and nuclei from k = 5 embryos. For A and B, light blue shading indicates GBE. Error envelopes indicate SD. (C) Still images showing cell deformation is required for nuclear deformation and dispersion indicated by fairly round nuclei present in Y-27632–injected (Y27) embryos compared to vehicle-injected (control) embryos. (D) Comparison of the major:minor axis percent change at nuclear midplane for cell (blue) and nucleus (red) between control and Y27 embryos. The negative percent change indicates a decrease in the axis ratio and the positive percent change indicates an increase in the axis ratio; n = 186 cell and nuclei for control and n = 236 cell and nuclei for Y27 from k = 3 embryos for each background. (E) Example of T1 transition with a color overlay of T1 interface (blue) shared by cells A and B, T2 vertex (yellow) shared by cells A–D, and T3 interface (red) shared by cells C and D (top). Bottom panels show the centroid (white) distance between AB (blue) and CD (red) cells. (F) Absolute nuclear midplane positions in AB, CD, and All cells. (G) Peak velocity with which nuclei move in apical–basal direction measured pre- (−5 to 0 min) and mid- (+15 to +20 min) GBE in wildtype embryos; n = 213 nuclei for pre- and n = 512 for mid-GBE from k = 3 embryos. (H) Maximum apical–basal speed of nuclei in AB, CD, and All cells. For F and H, n = 36 for AB and CD cells and n = 82 for All cells from k = 1 embryo. (I and I′) Comparison of probability change in relative nuclear position at pre- (−5 to 0 min, blue), early (0–10 min, yellow) and mid- (15–20 min, red) GBE in control and Y27 embryos; n = 3,339 nucleus pairs for control and n = 5,565 nucleus pairs for Y27 embryos from k = 3 embryos for each background. (J) Heatmap showing apical–basal distribution of nuclei (absolute midplane position) over the course of GBE in Y27 embryos. Color bar indicates the probability of nuclear midplane position; n = 1,097 nuclei from k = 3 embryos. (K) Maximum packing ratio achieved by clusters of nuclei as measured by the dispersion metric at early (0 min) and mid- (+20 min) GBE; for control, n = 370 clusters of cells for early and 336 clusters of cells for the mid-GBE, for Y27, n = 803 clusters of cells from early and 870 clusters of cells from mid-GBE from k = 3 embryos for each background. The measured n values are regardless of time points where not indicated. For C and E, scale bar = 10 and 5 μm, respectively. Statistical significance was calculated using the Mann–Whitney U-test. ns = not significant, ***P < 0.001, ****P < 0.0001.

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