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. 2001 Oct;12(10):2961–2974. doi: 10.1091/mbc.12.10.2961

Figure 1.

Figure 1

The subcellular localization of Cdc15-Ha during the cell cycle. Exponentially growing cells either lacking (A, no tag; K699) or carrying a CDC15-HA fusion (B, A1787) were fixed and subjected to indirect in situ immunofluorescence. Cdc15-Ha was visualized with the use of α-HA antibodies (BABCO). DAPI (4′6-diamidino-2-phenylindole) was used to stain DNA. (C) Cdc15-Ha localization in cells undergoing a synchronous cell cycle after an α-factor–induced G1 arrest. The percentage of cells with Cdc15-Ha on the SPB (spindle pole body) and the percentage of cells with metaphase and telophase spindles are shown. (D, E) nud1–44 (A2044) or tem1–3 (A2125) cells carrying a CDC15-HA fusion were arrested in G1 with α-factor (5 μg/ml) followed by release into medium lacking pheromone at the restrictive temperature (37°C). The percentage of cells with metaphase (closed circles) and anaphase spindles (closed squares) as well as the percentage of cells with Cdc15 on SPBs (open squares) or in the cytoplasm (open circles) was determined. (F) Wild-type (A1787) and bub2:: HIS3 cells (A2840) were arrested in early S phase with the use of hydroxyurea (10 mg/ml) followed by release into medium lacking the drug. The percentage of cells with metaphase (closed circles) and anaphase spindles (closed squares) as well as the percentage of cells with Cdc15 on SPBs (open squares) was determined at the indicated times. (G) Colocalization of Cdc15-Ha with the spindle pole body component Tub4 in bub2:: HIS3 cells. In all experiments, 200 cells were counted per time point (n = 200).