Fig. 4: Laminar wrinkling is sensitive to stiffness in three-dimensional (3D) culture and develops post mitosis.

a Confocal images of MDA-MB-231 cells expressing GFP-LMNA (green) stained with F-actin (magenta) in GelMA 3D culture. The scale bar is 10 μm. b Corresponding quantification of mean nuclear EFC ratio correlated to mean cell aspect ratio is shown for MDA-MB-231 cells expressing GFP-LMNA in GelMA 3D culture. n = 23 and 22 based on three replicates of soft (1–3 kPa) and stiff (10–15 kPa) gels, respectively. Error bars, SEM. c Confocal images of stained lamin A/C (green) and F-actin (magenta) in 344SQ lung cancer cells cultured on glass and acini in 3D Matrigel culture. The scale bar is 10 μm. d Corresponding quantification of nuclear EFC ratio is shown for 344SQ lung cancer cells cultured on glass and acini in a 3D Matrigel culture. n = 40 and 48 for 2D glass and 3D acini, respectively, based on three replicates. Error bars, SEM. p-values from the two-sided Mann–Whitney U-test. e Representative confocal images of lamin B1 (gray) stained in tumor microarray from n = 50 and 11 images of human invasive ductal carcinoma and larynx squamous cell carcinoma, respectively. The scale bar is 20 μm. f Time-lapse confocal images of a GFP-LMNA (gray) expressing fibrosarcoma cell and a fibroblast during cell mitosis. The scale bar is 10 μm. g Corresponding quantification of mean nuclear EFC ratio is shown for mother (red) and daughter cells (blue and green), normalized to the value in the first time-frame during cell mitosis. n = 4 and 15 for fibrosarcoma cells and fibroblasts, respectively, based on three replicates. Error bars, SEM.