FIGURE 5.

EDD siRNA reduces mitotic frequency and success rate, but not mitotic timing. A–C, asynchronous HeLa cells were treated with EDD, BUBR1, or scrambled control (Con) siRNAs for 12 h prior to time-lapse microscopy. Cells were scored for their ability to initiate (A) and successfully complete mitosis (B), and we measured the duration of those successful mitoses (C). D, SDS-PAGE and Western blotting with the indicated antibodies validated the efficacy of siRNA-mediated knockdown. E–H, the fate of cells attempting mitosis was also assessed for overall cell death (E), death during mitosis (F), death following mitosis (G), and abnormal cytokinesis (e.g. fusion of daughter cells) (H). The total number of cells analyzed was >100 and, for mitotic cells, >40 for all siRNA-treated cell groups. The data in A, B, and E–H were analyzed by Fisher's exact test with the p value indicated, whereas the data in C were analyzed using one-way analysis of variance and Tukey's multiple comparison test, which revealed a significant difference between BUBR1 siRNA and the other three siRNA treatments. E–H incorporate pooled data from siEDD-1 and siEDD-2. Errors bars represent mean ± S.D.