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. Author manuscript; available in PMC: 2009 Dec 1.
Published in final edited form as: FEBS J. 2008 Dec;275(23):5774–5784. doi: 10.1111/j.1742-4658.2008.06693.x

Figure 2. CDC14 is required for Clb2 degradation brought about by ectopic CDK inactivation.

Figure 2

GAL-SIC1 (A810), cdc14-3 GAL-SIC1 (A831) and cdc15-2 GAL-SIC1 (A844) cells were arrested with nocodazole (15µg/ml) for 165 minutes in YEP medium containing 2% raffinose at 23°C. Then the 0 time point was taken and cells were shifted to 37°C for 30 minutes. Then 2% galactose and α–factor (5 µg/ml) were added to induce Sic1 production and to inhibit Cln-CDKs, respectively. 5µg/ml nocodazole was readded at the same time to ensure that microtubules remain depolymerized. Samples were taken at the indicated times after temperature shift to determine the amount of Clb2 protein (A), Clb2-associated histone H1 kinase activity (B) and DNA content (C). Kar2 was used as a loading control in western blots. Methods were as described in Amon (1997).