Figure 5.

Microtubule detachment as seen in short time-lapse sequences in cnm67Δ cells. One astral microtubule bundle is formed at the SPB located in the bud and contacts the bud–cortex (0"–45"). Upon contact with the cortex the bundle rapidly changes its orientation (45" and 75"). Suddenly the bundle is no longer visible, although the spindle is perfectly in focus (90"). The microtubule bundle is detected again but does not show any connections to the SPB and is moving along the cortex (120"–225"). New microtubules at the SPB in the bud are formed (255"). Although the spindle is clearly captured by this z-image series, both SPBs are devoid of astral microtubules, indicating new detachment (495"). This is supported by two detached microtubule bundles that are localized in the bud and one in the mother cell. Bar, 5 μm. Movies 5 and 6: Microtubule detachment as seen in GFP-Tub1–labeled cnm67Δ cells followed for 14 and 22 min, respectively. Acquisition interval, 15 s; movie speed, 10 frames per second = 2.5 min/s. Three z-axis plane fluorescence images were acquired and merged.