FIGURE 6.

Knockdown of USP2a induces abnormal mitosis. 786-O cells on a 10-cm dish and on chamber slides were transfected with 20 nm control RNA, 20 nm USP2a siRNA#1, or 20 nm Aurora-A siRNA on day 1. The cells were transfected again on day 2 and were left in culture for another 2 days. A, the cells on the 10-cm dish were harvested, and Western blot analysis was carried out for Aurora-A and actin. B, the cells on chamber slides were fixed and stained for α-tubulin (green), γ-tubulin (red), and DNA (blue) as described under “Materials and Methods.” Mitotic index was determined. *, p < 0.05 (Student's t test). Error bars, S.D. C, typical examples of normal and abnormal mitotic cells. D, quantitative assessment of normal and abnormal mitotic spindles. For each treatment condition, >150 mitotic cells were scored. Data are representative of three independent experiments. *, p < 0.05 (Student's t test).