Perturbation of passive nuclear transport dynamics alters nuclear bleb formation. (a) Percentage of HT-1080 cells displaying nuclear blebbing on the front or rear of the nucleus after being exposed to a hypotonic shock (165 or 85.5 mOsm) at their leading edge (n ≥ 3 independent experiments with a minimum of 20 cells per experiment). (b) t_1/2 of PA-GFP signal diffusing from the nucleus to the cytoplasm of VC or LMB-treated (20 nM) HT-1080 cells plated on 2D (n ≥ 29 cells from three or more independent experiments). (c) Percentage of VC and LMB-treated (20 nM) HT-1080 cells in confinement and on 2D displaying nuclear blebbing, as observed from cells fixed and stained with Hoechst (n ≥ 3 independent experiments with a minimum of 20 cells per experiment). (d) Percentage of cells displaying nuclear blebs after treatment with vehicle, Y27632, LMB, or Y27632 and LMB (n ≥ 5 independent experiments with a minimum of 14 cells per experiment). (e) t_1/2 of PA-GFP signal diffusing from the nucleus to the cytoplasm of scramble control or TPR-knockdown HT-1080 cells plated on 2D (n ≥ 29 cells from three or more independent experiments). (f) Percentage of scramble control and TPR-knockdown HT-1080 cells in confinement and on 2D displaying nuclear blebbing, as observed from cells fixed and stained with Hoechst (n ≥ 3 independent experiments with a minimum of 20 cells per experiment). Values represent mean ± SD (b and e) or mean ± SEM (a, c, d, and f). *, P < 0.05; **, P < 0.01 relative to vehicle/scramble control or isotonic rear bleb; §§, P < 0.01 relative to Y27632.