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. 2006 Jan 30;172(3):335–346. doi: 10.1083/jcb.200507162

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Copyright © 2006, The Rockefeller University Press

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Figure 2. — Expression of Bub2-myc9 delays mitotic exit in cdc5-2 cells at permissive temperature. Cultures of wild-type (ySP41), MET3-CDC5 cdc5-2 (ySP3426), and MET3-CDC5 cdc5-2 BUB2-myc9 cells (ySP3480), exponentially growing in –Met medium, were arrested in G1 with α factor and released at 25°C in YEPD supplemented with 2 mM Met (MET3-CDC5 expression off). After 120 min, 10 μg/ml α factor was readded to prevent cells from entering a second cell cycle. Cells were collected at the indicated times for FACS analysis of DNA contents (A); kinetics of budding, nuclear division, and spindle elongation/breakdown after in situ immunostaining of tubulin (B); and Western analysis of Clb2 and Sic1, as well as Clb2/Cdk1 immunoprecipitations followed by kinase assays using histone H1 as substrate (C). Swi6 was used as loading control.