FIG. 3.

(A) Cdc14-GFP is not prematurely dispersed in clb5Δ cells. Wild-type (left panels) and clb5Δ (right panels) samples at various times are shown. The graphs depict the extents of nuclear division and Cdc14-GFP dispersion. DAPI, 4′,6′-diamidino-2-phenylindole. (B) Insufficient phosphorylation of Hct1 in a clb5Δ strain. Wild-type (US2352) and clb5Δ (US2354) cells carrying GAL-HA₃-HCT1 were arrested in G₁ by α-factor treatment in YEP+Raff+Gal at 24°C. The cultures were filtered, washed, and resuspended in YEP+Glu at 24°C. Samples were analyzed for DNA content, nuclear division index, Cdc28 protein levels, and Hct1 phosphorylation. (C) Inefficient phosphorylation of Hct1 in clb3Δ clb4Δ clb5Δ cells. clb3Δ clb4Δ clb5Δ cells carrying MET3-HA₃-HCT1 on a 2μm plasmid (US2438) were synchronized in G₁ by α-factor treatment for 3 h (the first 1.5 h in YEP+Raff+Met and the second 1.5 h in −Met+Raff medium). The culture was divided into two halves; one-half of the cells were resuspended in YEP+Raff+Met, and the other half were resuspended in YEP+Gal+Met. Samples were analyzed for DNA content, Hct1 phosphorylation, and Clb2 and Cdc28 protein levels.