Enhanced dispersion of nuclei in the absence of the deformation pathway. (A) Heatmap showing the apical–basal distribution of nuclei (absolute midplane position) for control (luciferase shRNA) and kuk shRNA embryos. Color bar indicates the probability of nuclear midplane position; n = 888 for control and n = 929 nuclei from k = 3 embryos for each background. (B) Probability distribution of relative depth of nuclei at pre- (−5 to 0 min, blue), early (0–10 min, yellow), and mid- (15–20 min, red) GBE for control (luciferase) and kuk embryos; n = 21,303 pairs of nuclei for control (luciferase) and n = 13,857 pairs of nuclei for kuk from k = 3 embryos for each background. (C) Maximum packing ratio of nuclear clusters at pre- (−5 min) and mid- (+20 min) GBE for kuk embryos; n = 580 clusters of cells from k = 3 embryos. (D) Nuclear position (nucleus area, blue) correlates with cell bulging (indicated by increased cell area, red). Cell deformation is enhanced by the deformation-resistant nuclei (kuk). Negative x-axis values indicate apical regions above nuclear midplane and the positive values indicate the basal regions; n = 317 control and n = 357 kuk cell-nuclei pairs from k = 3 embryos for each background. Error envelopes indicate SD. (E) Side views of 3D cell reconstruction from time-lapse movie of control (luciferase) and kuk embryos depicting higher degree of cell deformation in cells with non-deformable nuclei (kuk). The measured n values are regardless of timepoints. For C, statistical significance was calculated using the Mann–Whitney U-test. *P < 0.05.