FIG. 5.

Cdc13p is degraded in APC cut mutants. (A to C) Wild-type (A), lid1-6 (B), and cut9-665 (C) cells were grown to mid-log phase at 25°C in YE medium, synchronized in early G₂ phase by centrifugal elutriation, and then shifted to 36°C. Cells were collected at 15-min (for wild-type cells) or 20-min (for APC mutants) intervals, and the septation index and percentage of cut cells were determined in each sample. Protein lysates were also prepared from each sample. Cdc2p and Cdc13p levels were determined by immunoblotting with PSTAIR monoclonal antibodies and affinity-purified anti-Cdc13p antibodies, respectively. (D) The DNA content of each sample was also determined by flow cytometry.