Fig. 2

ENSA depletion delays mitotic entry and impairs mitotic spindle pole integrity. A Fucci-HeLa cells were transfected with the control or ENSA siRNA. Time-lapse images from 48 h after transfection are shown. The duration from nuclear envelope breakdown (entry initiation) to the formation of the rounded-up shape (entry completion) was determined for the control and ENSA-depleted cells. Images were captured every 3 min to monitor mitotic progression. B Quantification of mitotic entry in the control and ENSA-depleted HeLa cells (n = 50 cells). Scale bar, 5 μm. Error bar, SEM. C-H HeLa cells were transfected with the control or ENSA siRNA for 48 h. Quantification of mitotic cells is shown (C). Cells were stained with the indicated antibodies (D). Scale bar, 5 μm. E The percentage of metaphase cells with misaligned chromosomes among the total metaphase cells. F Quantification of spindle pole abnormalities during mitosis. G Quantification of spindle abnormalities during mitosis. Data are presented as the mean ± SEM of values from at least three independent experiments (n = 50 miotic cells for each quantification and group). ***p < 0.0001. H Cells were stained with the indicated antibodies (left panel). Scale bar, 5 μm. Quantification of centrosome defects during mitosis (right panel)