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Figure 2


Fig. 2. (A,B) Cell cycle progression of cells expressing mutant forms of CLB2. The indicated cells were synchronously released from an {alpha}-factor-induced G1 arrest at 30°C. Aliquots were taken every 15 minutes. (A) DNA content was analyzed by flow cytometry and (B) the percentage of budded (crosses), pre-anaphase (triangles) and binucleate (squares) cells determined microscopically after a brief sonication. Pre-anaphase cells were defined as having the nucleus located at the bud neck. Note that in our strain background, about 20% of clb2{Delta} cells had the nucleus at the bud side of the neck just before anaphase and were classified as pre-anaphase cells. Each panel is representative of at least two independent experiments. More than 200 cells were counted for each time point. (C) Cells from the strain W303 clb1,3,4{Delta} clb2ts (Amon et al., 1993) were transformed with the indicated vectors and assayed for growth at 36°C. Serial 5x dilutions of saturated cultures were spotted on YPD plates and incubated at the indicated temperatures for 36 hours.





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