Figure 4. Overexpression of LMW-E overcomes nocodazole-dependent mitotic arrest.
A, HeLa H2B-GFP cells were infected with LacZ-control, EL, or LMW-E expressing adenovirus and then treated with nocodazole (Noc, 100 ng/ml) for 16 h. Cyclin E expression was detected by western blot analysis. B, Mitotic cells treated as in A were counted by microscopy and the mitotic index was calculated +/- S,D. (N=1000 for two independent experiments). C, Human fibroblast MRC50 cells were infected with adenovirus expressing LacZ control, EL, or LMW-E and then exposed to nocodazole for the indicated times and analyzed by flow cytometry. The graphs show the percentages of cells in the G0/G1, S, and G2/M phases and are representative of three independent experiments. D, Western blot analysis of cyclin E, cyclin B1, and vinculin expression in cells treated as described in C. Densitometry was performed using ImageQuant software with the cyclin B1:vinculin ratio.