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Figure 4 | BMC Cell Biology

Figure 4

From: Human ASPM participates in spindle organisation, spindle orientation and cytokinesis

Figure 4

siRNA mediated ASPM induces misorientation of the plane of cell division, cytokinesis failure and increased apoptosis in U2OS cells. A. Panels showing phase contrast images of living U2OS cells taken from time-lapse movies. Time points are shown in the bottom left hand corner of each image. Top row. Normal symmetrical division of a GL3 siRNA treated control cell with the cleavage plane oriented perpendicular to the cell substrate (see Additional file 5). Scale bar = 10 μm. Middle row. ASP1 siRNA treated cell undergoing an asymmetric division with the mitotic spindle positioned perpendicular to the cell substrate. This division led to cytokinesis failure with the formation of a binucleate cell. Arrowhead shows the upper daughter cell moving out of the focal plane towards the observer as the cleavage furrow is formed parallel to the surface of the imaging dish. An arrow indicates the lower daughter cell still attached to the surface of the dish (see Additional file 6). Scale bar = 10 μm. Bottom row. Binucleated ASP1 siRNA treated cell undergoing apoptosis (see Additional file 7). Scale bar = 10 μm. B. Flow cytometric analysis of caspase-3 assayed 96 hours after ASP1 and GL3 siRNA treatment. An increase in the population of apoptotic (dead) cells is observed in ASP1 siRNA experiments. C. Histogram showing the mean percentage of apoptotic cells in ASP1 and GL3 siRNA caspase-3 assays. A 2.7 fold increase in the percentage of apoptotic cells in the ASP1 siRNA population was identified compared to the GL3 siRNA population. Standard deviation values for the mean are shown as vertical bars.

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