Fig. 3: Loss of endogenous Maspin promotes cytoskeleton rearrangements in epithelial cells.

a Maximum intensity projections for MCF-10A KO versus MCF-10A WT cells during interphase co-stained for F-actin and α-tubulin. Insets represent reduced cortical F-actin for KO as compared to WT cells. Scale bar: 10 µm; (b) Cortical F-actin fluorescence intensity for MCF-10A WT (n = 59), KO C10 (n = 41 cells) and KO D9 (n = 56 cells); c Live imaging of MCF-10A WT versus KO cells upon mitotic entry. Black arrows indicate retraction tails. Scale bar: 25 µm; (d) Cell area prior to mitotic rounding WT (n = 49 cells), KO C10 (n = 49 cells), KO D9 (n = 49 cells); (e) Maximal tail length for WT (n = 50 cells), KO C10 (n = 48 cells) and KO D9 (n = 50 cells) cells during mitotic progression; (f) Mitotic duration for WT (n = 50 cells), KO C10 (n = 48 cells) and KO D9 (n = 50 cells). Cells were imaged at interval frames of 5 min; (g) MCF-10A cells released after nocodazole blockage and co-stained for F-actin, α-tubulin and nuclei. Heatmaps indicate distribution of F-actin fluorescence intensity in both MCF-10A WT and KO cells. Scale bar: 10 µm and 5 µm (inset); (h) Cell roundness was measured for MCF-10A WT (n = 59 cells), KO C10 (n = 53 cells) and KO D9 (n = 53 cells) at metaphase; (i) Representation of mitotic measurements (d1 and d2); j Spindle length (d2) was measured for WT (n = 43 cells), KO C10 (n = 48 cells) and KO D9 (n = 58 cells); (k) Cortical distance along spindle axis (d1) measured for WT (n = 43 cells), KO C10 (n = 52 cells), KO D9 (n = 52 cells) cells at metaphase; l Cortical distance along spindle axis to spindle length ratio (d1/d2) measured for WT (n = 43 cells), KO C10 (n = 52 cells) and KO D9 (n = 50 cells) at metaphase. n.s. = not significant.