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. 2003 Nov;14(11):4486–4498. doi: 10.1091/mbc.E03-02-0095

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Figure 6. — The ectopic release of Cdc14 from the nucleolus brought about by GAL-CDC5 and deletion of BUB2 depends on TEM1. (A, B) GAL-CDC5 bub2Δ (A4453) and GAL-CDC5 bub2Δ tem1-3 (A4549) cells carrying a CDC14-3HA and a CFI1-3MYC fusion were grown and analyzed as described in Figure 5 except, cells were shifted to 37°C at the time of galactose addition. (C, D) GAL-CDC5 (A4450) and GAL-CDC5 tem1-3 (A4547) cells all carrying a CDC14-3HA and a CFI1-3MYC fusion were arrested in early mitosis with the microtubule depolymerazing drug nocodazole (15 μg/ml) in YEPR at 25°C. After 2.5 h, cells were shifted to 37°C in the presence of nocodazole. Samples were taken at the indicated times to analyze the percentage of cells with Cdc14 released from the nucleolus (C) and Cfi1/Net1, Cdc14 and Cdc5 protein levels and mobility in SDS-PAGE (D). (E) GAL-CDC5 bub2Δ (A4453) and GAL-CDC5 bub2Δ esp1-1 (A4282) cells carrying a CDC14-3HA and a CFI1-3MYC fusion were arrested with 10 mg/ml HU at 25°C. Galactose was then added and cells were shifted to 37°C and Cdc14 localization was analyzed at the indicated times.