Fig. 3. Overexpression of pin1+ causes a severe growth defect. (A) Wild-type (Sp13) and overexpression (Sp15) strains were grown on plates containing thiamine for two days and inoculated into non-repressive medium for 20 hours, which is the time required for induction of pin1+ overexpression. Cells with overexpressed pin1+ were re-inoculated into non-repressive medium to OD595=0.05 and cultured at 32°C. For every 12 hours, aliquots of cells were taken and the cell densities (OD595) were measured. (B) Same as A except that cell aliquots were fixed in 1 ml of cold 100% ethanol, resuspended in 50 mM sodium citrate and stained with 1 µM Sytox Green. Flow cytometry was performed with a Becton Dickinson FACScan and data were analyzed by CellQuest software.
