SPB settling occurs regardless of bouquet status. (A and B) Frames from films of meiocytes with endogenously GFP-tagged Hrs1 and nmt1-controlled Ish1-GFP (to visualize the NM) along with the indicated markers (as in Fig. 2). Bars: (black) 5 µm; (gray) 1 µm. (A) In WT meiocytes, the SPB colocalizes with the NM throughout meiosis. (B) In bqt1Δ meiocytes, the SPB detaches from the NM only after SPB movement ceases, i.e., upon disappearance of Hrs1-GFP (t = 0′). (C) Quantitation and timing of phenotypes shown in A and B from three independent experiments analyzing 30 cells for each strain. Note that dislodgement of the SPB from the NM is never observed in WT meiocytes. SPBs of bqt1Δ cells dissociate from the NM only after Hrs1 signal has disappeared. (D and E) Kymographs of Hht1-CFP (Chromatin) and Sid4-mCherry (SPB) during prophase, MI, and MII. The SPB settling phase (yellow) is shown magnified. The data shown are from a single representative experiment out of >10 repeats. (F) Time (in minutes) for which SPB (as visualized via Hrs1-GFP) settles after prophase. n is the total number of cells scored from greater than six independent experiments. Error bars show the mean SD.