(A) Wild-type cells (HPH164) were grown in YPDS medium at 30°C. Microtubules and DNA are visualized by GFP-Tub1 and Hht1-mCherry fluorescence, respectively. cMTs are marked by yellow arrows. a, b: unbudded G1 cell; c, d: preanaphase cell with monopolar nuclear MTs; e, f: preanaphase cell with bipolar spindle; g: anaphase cell. Scale bar, 2 µm. (B) Quantification of (A). G1, S/G2, G2/M, and anaphase represent unbudded cells, small budded cells with single unduplicated SPB, medium budded cells with 2 SPB, and large budded cells with an SPB in both the mother and the bud, and large budded cells with elongated spindle, respectively. n > 50 cells for each category. Ana, anaphase. (C) Cells containing the cMT detached from the SPB in (A). Scale bar, 2 µm. (D) Analysis of the duration of cMT persistence and cMT re-establishment time at SPBs in preanaphase cells with a bipolar spindle by time-lapse microscopy. GFP-TUB1 cells (HPH194) were grown in SD complete medium at 30°C. Consecutive sections were taken every 30 s. Duration of continuous cMT presence was scored as cMT persistance, while the time between the loss of cMT and the acquisition of a new cMT was scored as cMT re-establishment. Total recording time was 38460 s.