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Fig. 5. Septins disassembled from the neck assemble into new rings at the bud tip during G2 but not M phase. (A) Strain JMY1141 (cdc12-6) was grown to exponential phase in YEPD at 24°C, shifted to 37°C for 30 min, and then shifted back down to 24°C. Samples were taken before the shift (24°C), 30 minutes after shift-up (
), and at various times after shift-down (
). Arrow indicates original neck. For each sample following shift-down, we quantitated the percentage of cells with septins assembled at the tip or within the bud (B) or as rings in unbudded cells or at the neck (C). These categories were separated in the interests of clarity, and the remaining cells (25%, not graphed) showed no septin staining. (D) Cells were arrested in mitosis (75% large budded cells) by treatment with 15 µg/ml nocodazole for 2 hours at 24°C and then subjected to the same shift-up/shift-down regimen as above. Bar, 10 µm.
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