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Fig. 3. cdk5^ts cells are affected in polarity. (A) Calcofluor staining of cdk5^ts cells grown in liquid CMD medium for different times at the restrictive temperature (34°C). Note that upon temperature shift, cdk5^ts cells expand their diameter and lose their bud constriction, giving cells separated by septation. Bar, 20 µm. (B) Defects in polar growth during G2 arrested cdk5^ts cells. Cells overexpressing wee1 arrest at G2 producing elongated buds. However, in cdk5^ts cells there is no bud formation and instead two areas of isotropic growth at the poles are apparent. Inset, DAPI staining showing that in cdk5^ts cells there is only one nucleus in agreement with a G2 cell cycle arrest. Bars, 10 µm. (C) Effect of cytoskeletal inhibitors on plate growth of wild-type and cdk5^ts strains. No difference between cdk5^ts and wild-type cells was found in the presence of the MT-destabilizing drug benomyl (1 µM). By contrast, cdk5^ts cells are more sensitive to the actin inhibitor cytochalasin D (CytoD; 10 µM).

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