FIG. 5.

Combined effect of nocodazole with UCN-01 or caffeine on SV40-infected cells. Confluent CV-1 cells were infected with SV40 at 100 PFU per cell and treated with mimosine as described in the legend to Fig. 1. At 3 and 6 h after release from mimosine, the cells were incubated with 6 mM caffeine and 300 nM UCN-01, respectively, in the presence or absence of nocodazole (0.1 μg/ml). Samples were harvested at 48 hpi. (A) Expression of P-H3 and change in chromosome morphology. The number of P-H3-positive cells was also determined. Normal mitotic condensation, black; ACC, light gray; P-H3, dark gray. (B) Percentage of floating cells obtained from each of the treatments described above. (C) Whole-cell lysates were resolved by SDS-PAGE and immunoblotted. Each filter was reacted with antibodies specific to each protein and then with alkaline phosphatase-conjugated goat anti-mouse or -rabbit antibody. Exp, experiment.