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Fig. 6. Aberrant Cnk2p levels affect cell cycle progression. (A) Mitotic index measurements of wild-type (WT), CNK2-HA, CNK2-RNAi and fa2-1 cultures. Cultures were synchronized by alternating light-dark cycles for 3 days and examined microscopically on day 4 for the percentage of cells with cleavage furrows. A total of 200 cells were examined for each time-point, from two independent experiments (100/experiment). (B) Growth of WT, CNK2-HA and CNK2-RNAi cells. Cultures were grown in minimal media for several days in constant light and placed in the dark for 24 hours. Cultures were shifted back to the light to initiate growth and samples were taken every 2 hours. A total of 200 cells were measured for each time-point from two independent experiments (100/experiment), error bars represent s.e.m. (C) Analysis of the number of division cycles post commitment. Cultures were grown in TAP media for several days, plated on minimal media, and placed in the dark for 24 hours. Colonies were examined microscopically to determine the number of cells per colony. A total of 600 colonies were scored from two independent experiments (300/experiment). (D) Commitment size measurement. Cultures were grown as in B, and a sample was fixed immediately after the shift to light. A total of 300 cells were measured from each culture in three independent experiments (100/experiment).
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