Supplementary Figure 7: Myosin IIA depleted mitotic cells are compromised in sustaining persistent pressure.
From: Cdk1-dependent mitotic enrichment of cortical myosin II promotes cell rounding against confinement
(a) Representative confocal images of STC-arrested mitotic MYH9-GFPLifeact-mCherry expressing HeLa cells confined to a height of 10 μm. Cells were transfected at least 48 hours prior to experiments with control (n = 9 cells), Ect2 (n = 13 cells) or MYH9MYH9 (n = 10 cells) siRNA. Scale bars, 10 μm. (b) Cortex/cytoplasm ratios of F-actin (red) and myosin II (green). Cells were transfected at least 48 hours prior to experiments with control (n = 9 cells), Ect2 (n = 13 cells) or MYH9 (n = 10 cells) siRNA. (c) Average force traces of STC-arrested mitotic MYH9-GFP Lifeact-mCherry expressing HeLa cells confined to a height of 10 μm. Time zero represents arrival at the height of 10 μm after the AFM-cantilever descended at a speed of 1 μm s−1. Cells were transfected with control (orange, n = 9 cells), Ect2 (red, n = 13 cells) or MYH9 (purple, n = 10 cells) RNAi. (d) Transient or persistent pressure (grey) and tension (blue) of mitotic cells under confinement. Cells were transfected at least 48 hours prior to experiments with control (n = 9 cells), Ect2 (n = 13 cells) or MYH9 (n = 10 cells) siRNA. The diamond-box contains 25–75% percentiles of the data and the bar denotes the median. Mann–Whitney significance tests of mitotic cells with respect to data plotted in the first column are indicated: (NS) U > 0.05, (∗) 0.05 ≥ U > 0.005, (∗∗) U ≤ 0.005.
