Fig. 3. iASPP is required for the completion of cytokinesis.

a Depletion of iASPP or CEP55 by siRNAs in HeLa cells. HeLa cells were transfected with indicated siRNAs. After 48 h, cell lysates were prepared for WB with indicated antibodies. b FACS analyses of the DNA content of cells depleted of iASPP, CEP55, or treated with control siRNAs. Peaks corresponding to 2n, 4n, and 8n DNA contents are labeled, and the cell percentage volumes are provided. c HeLa cells transfected with the indicated siRNAs were stained with β-tubulin antibodies (red) and DAPI (blue). d, e Quantification of multinucleated cells (d) and cells arrested at the midbody (e) in control, iASPP-depleted, or CEP55-depleted HeLa cells. All data shown are mean values ± SD (error bar) from three replicates. **p < 0.01, Student’s t-test. f Representative time-lapse phase-contrast images of a control cell proceeding through mitosis is shown (upper panel). In the lower panels, representative time-lapse images of iASPP-depleted cells with either one or two nuclei proceeding through mitosis are shown. Black arrowheads highlight cells that are starting to divide. g Quantitation of post-metaphase HeLa cells treated with control or iASPP siRNAs. All data shown are mean values ± SD (error bar) from three replicates. **p < 0.01, Student’s t-test. h iASPP depletion leads to increased cell death. HeLa cells were transfected with control or iASPP siRNAs. After 72 h, the cell death was measured by flow cytometry using the propidium iodide staining assay. All data shown are mean values ± SD (error bar) from three replicates. **p < 0.01, Student’s t-test