Fig. 6. Cdc5 controls Bfa1 and Lte1 phosphorylation. (A) Strain SLY109 (3HA-BFA1 cdc5^ts (msd2-1)) was grown to mid-log phase at 25 °C and sampled. Some of the remaining cells were transferred to 37°C and nocodazole was added to others with incubation at either 25°C or 37°C. Following protein extraction an immunoblot was prepared and probed with 12CA5. (B) Strain SLY109 was synchronised with -factor and released at either 25°C or 37°C. Samples were removed at the times indicated and an immunoblot prepared. The lower panel shows budding curves for the two cultures. (C) Mid-log cells of strain SLY109 were treated with nocodazole for 3 hours at 25°C. Incubation of one half of the culture was continued at this temperature, whereas the other half was transferred to 37°C. After sampling, an immunoblot was prepared from the cells. (D) Strains SJY121 and 122 were grown to mid-log phase at 25°C. The cultures were split and one half was incubated with nocodazole. Following 3 hours incubation at 30°C, cells were harvested and an immunoblot prepared from the extract. Note that longer exposure did not reveal further phosphorylations in lanes 1 and 2.