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. 2008 Mar 10;180(5):931–945. doi: 10.1083/jcb.200711014

Figure 1.

Figure 1.

mih1Δ cells have an increased cell size and undergo delayed entry into mitosis. (A) Cells of the indicated genotypes were grown to log phase in YPD media at room temperature. Cell size was measured using a Z2 Coulter counter. (B) Wild-type and mih1Δ cells were synchronized with α-factor and released into fresh YPD media and samples were taken at 10-min intervals. Cells were fixed and stained with an anti-tubulin antibody and the percentage of cells with short spindles was determined. Over 200 cells were analyzed for each time point. The experiment was repeated at least three times with matched controls performed in parallel on the same day, and the same overall trend was observed. Error bars are not displayed because the timing of release from α-factor arrest varies from day to day. (C) Wild-type and mih1Δ cells were synchronized with α-factor and released into fresh YPD media and samples were taken at 10-min intervals. Cells were fixed and the percentage of cells with small buds was determined. Over 200 cells were analyzed for each time point. The experiment was repeated at least three times with matched controls performed in parallel on the same day, and the same overall trend was observed. Error bars are not displayed because the timing of release from α-factor arrest varies from day to day. (D) Wild-type and mih1Δ cells carrying CLB5-3XHA cells were synchronized with α-factor and released into fresh YPD media and samples were taken at 10-min intervals. Western blotting was used to monitor the behavior of Clb5-3XHA. (E) Wild-type cells were synchronized with α-factor and released into fresh YPD media and samples were taken at 10-min intervals. Western blotting was used to monitor the behavior of Swe1, Clb2, and Cdk1 tyrosine 19 phosphorylation. The exposures of the Western blots were normalized using background bands to allow direct comparison of the signals from each strain. The Swe1 protein migrates as a disperse group of phosphorylated forms above a 97 kD size standard, whereas Clb2 and Cdk1 migrate approximately at 50 and 35 kD, respectively.