Fig. 3. The effects of cdc2-as on meiosis, and rescue by increased expression of cdc13. (A) A cross of h⁺ cdc2-as with h^- cdc2-as lys1-131 was performed at 25°C. After 2 days, the mating mixture was suspended in PBS, fixed, and stained with DAPI and Calcofluor. The outcomes of the mating are indicated on the histogram. The figures indicate the percentage of each outcome class; more than 300 spore-containing cells were scored. The combined DAPI and transmission image of a dyad is shown. (B) Small cells were selected from an exponentially growing cdc2-as culture by centrifugal elutriation, and inoculated into fresh medium in the presence of 10 µM analogue or DMSO and incubated for 3 hours at 36°C. Cells were fixed and stained with DAPI and Calcofluor. (C) cdc2-as cells were transformed with a plasmid containing the genomic copy of cdc13 or empty vector. Colonies were allowed to form on EMM2 medium at 25°C, replicated to 36°C and incubated at 36°C overnight. Colonies were photographed. The inset shows a magnification of the edge of the colony. (D) The cells described in panel C were treated with the indicated concentrations of analogue at 25°C and samples were analysed by FACS. Scale bars: 10 µm.