Fig. 3
Depletion of mitotic cell mechanics genes is linked to impaired cortical targeting of myosin II. a Depiction of the assay to measure rounding pressure of mitotic cells. STC-arrested mitotic cells were confined to a set height of 12 µm by a microcantilever equipped with a terminal wedge. Rounding pressure P was calculated from force F and contact area derived from images at the cross-sectional midplane (Methods). Cortex to cytoplasm ratio of fluorescence markers for myosin II (MLR myosin localization ratio) and actin (ALR actin localization ratio) were calculated as described in Methods11. b Relative equilibrium rounding pressure of STC-arrested confined mitotic cells depleted of the indicated proteins by RNAi. Blue dashed line, average control rounding pressure, normalized to one. Each black diamond represents one cell. ‘(n)’ denotes number of cells measured. Blue bars, mean. Statistical significance was determined by Mann–Whitney U-test (*p ≤ 0.05; ***p < 0.001). c Representative fluorescent confocal images of STC-arrested confined mitotic cells expressing markers for myosin II (MYH9-EGFP) and F-actin (Lifeact-mCherry), depleted of selected proteins as indicated. MLR and ALR, relative to control cells, are shown at the top right of each image. Scale bars, 10 µm. See Supplementary Fig. 5 for full results of rounding pressure and MLR/ALR for all genes tested
