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. 1998 Oct 5;143(1):135–145. doi: 10.1083/jcb.143.1.135

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Time-lapse series of spindle behavior in cdc28-4 or cdc28-4 clb5 diploids. Cells were grown in synthetic galactose medium followed by transfer to synthetic dextrose medium for 6 h at 23°C before mounting on the same medium for microscopy as described in Materials and Methods. Numbers correspond to the time elapsed in minutes. Representative series of the mutant (A– B) and parental (C) strains are shown. (A) The cell to the right contains a spindle positioned into the bud that underwent limited elongation. The cell to the left proceeded through anaphase. Selected frames from a 40-min time lapse experiment are shown. Bar, 5 μm. (B) Astral microtubular defects associated with spindle positioning and anaphase progression in cdc28-4 clb5 GAL1: CLB5 diploids. Cells improved alignment along the mother– daughter axis, but failed to restore position at the bud neck. Astral microtubules directed towards the mother cortex lengthened and curved. By an additional 20-min period after the last time point shown, cells had not proceeded through anaphase. Selected frames from a 40-min time lapse experiment are shown. Notice that the cell on the upper right side assembled a spindle throughout the time lapse. Bar, 5 μm. (C) Spindle positioning in a cdc28-4 diploid (MYT1010). Selected frames from a 20-min time lapse experiment to show parental behavior of astral microtubules and spindle positioning. Bar, 5 μm.