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Figure 9


Fig. 9. cdc14{Delta} cells show upregulated levels of B-cyclins. (A) Protein extracts from asynchronous cultures of the indicated strains grown in YPD were prepared and analyzed by western blot using 12CA5 antibodies to detect HA-tagged proteins. Triangle, Clbs-HA; diamond, a cross-reacting band used as loading control. (B) Flow cytometry analysis of asynchronous cultures of strains CDC14 and cdc14{Delta} grown in YPD or SC media at 30°C. (C) G1 cells from strains JC64 (CDC14/CDC14; CLB2HA/CLB2) and JC70 (cdc14{Delta}/cdc14{Delta}; CLB2HA/CLB2) were isolated by elutriation and released into YPD at 30°C. Samples were collected at intervals of 15 minutes after release, and protein extracts were prepared and assayed by western blotting for Clb2p-HA levels. Cdc28p (anti-PSTAIRE antibody) was used as loading control. Samples were also monitored for bud morphology and DAPI staining.





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