Cells expressing Pcp1–Ppc89–GFP undergo normal mitotic growth, and forced Cut12–SPB association has no effect on SPB quantity control. (A) Representative time-lapse images of zygotes expressing Pcp1-GFP (SPB) and mCherry-Atb2 (microtubules; upper panels) or Pcp1–Ppc89–GFP (SPB) and mCherry-Atb2 (microtubules; lower panels). Arrowhead indicates a multipolar spindle with supernumerary SPBs formed at MI. Time 0 is an arbitrary time point in prophase. Scale bars, 5 µm. (B) Microscopic images of mitosis in cells expressing Pcp1-GFP (SPB) and mCherry-Atb2 (microtubules; left panels) or Pcp1–Ppc89–GFP (SPB) and mCherry-Atb2 (microtubules; right panels). Scale bar, 3 µm. (C) Serial dilution growth assays for each indicated strain. (D) Schematic of Cut12–Ppc89–GFP construct used to force association between Cut12 and SPB (left). Quantification of asci containing extra spores in the indicated crosses (right). Cut12, Cut12-GFP; Cut12-Ppc89, Cut12–Ppc89–GFP. Error bars are SDs from three independent experiments (n ≥ 200 cells each; one-way ANOVA). (E) Representative microscopic images of prophase and MI in zygotes expressing Cut12–Ppc89–GFP (SPB) and Hht1-mCherry (chromatins). Scale bar, 5 µm. (F) Representative microscopic images of azygotic and zygotic asci expressing Plo1–Sid4C–GFP or Sid4C-GFP. Plo1–Sid4C–GFP fusion construct is used to tether Plo1 to SPB during meiotic prophase. Percentages of asci with extra spores are shown in the lower right. Scale bar, 5 µm. DIC, differential interference contrast.