Fig. 3. Prc1 is required for timely spindle bipolarization in MI.
a Live imaging of oocytes after Prc1 RNAi. Ndc80f/f oocytes labeled with EGFP-Map4 (microtubules, green) and H2B-mCherry (chromosomes, magenta) were used. Spindle shapes were reconstructed in 3D. Four independent experiments were performed. Scale bar, 10 μm. See also Supplementary Movie 6. b Prc1 depletion delays spindle bipolarization. Spindle shapes in 3D were categorized based on the aspect ratio, surface irregularity, and stability (see Methods). The plot shows the time at which a stable bipolar spindle was established (n = 32, 29 oocytes from four independent experiments). Mean +/− SD are presented. ****p < 0.0001 (p = 2.4E−08) by two-tailed unpaired Student’s t-test. c Prc1 depletion increases chromosome misalignment. Oocytes were categorized into ‘aligned’ when all chromosomes located at the middle half of the spindle (n = 32, 31 oocytes from four independent experiments). d Prc1 depletion causes a delay in anaphase I onset. Oocytes were incubated with or without the checkpoint inhibitor reversine (rev). The percentages of oocytes that underwent anaphase I onset were plotted (n = 32, 31, 14, 14 oocytes from at least three independent experiments).