Ndel15A expression prevents accumulation of Lis1 and DIC at the NE in response to nocodazole. (A) 10-μM nocodazole delays NEBD in COS-7 cells and results in an increase in cells with Lis1 (green) at the NE. Arrow, prophase cell; arrowhead, G2 cell. (B) Ndel1 (top left) and DIC (top right) accumulate at the NE (red arrows) in a nocodazole-treated cell. Ndel15A (bottom left) but not DIC (bottom right) accumulates at the NE (white arrows) in a nocodazole-treated cell. Ndel1 constructs are myc tagged. (C) Lis1 at the NE was reduced by Ndel15A (bottom right). White arrows indicate accumulation of myc-tagged Ndel1 or Ndel15A at the NE. Blue arrows indicate the accumulation of Lis1 at the NE. (D–F) NE accumulation of Ndel1, DIC, and Lis1 was quantified (see Materials and methods). Statistical significance was determined by Student's t test. (G) COS-7 cells were cotransfected with Ndel1, cdk5, p25, HA-cdk1, and cyclin B (Ndel1) or Ndel15A, dncdk5, and HA-dncdk1 (Ndel15A) and exposed to nocodazole for 3 h. Nuclear and cytosolic extracts were examined by Western blotting. Error bars indicate ±SD. Bars, 10 μm.