Supplementary Figure 3: Bouquet deficient meiocytes fail to properly assemble outer kinetochores.

(A–C) Nnf1 and chromosomes are observed via endogenously tagged functional Nnf1-GFP and Hht1-mRFP (as in Fig. 1), respectively. Nnf1 disappears from centromeres in early prophase and relocalizes to centromeres 10-40 min before metaphase I. Labels as in Supplementary Fig. 1. (A) wt meiosis. Nnf1-GFP appears on all chromatin masses at MI and MII. (B,C) bqt1Δ meiosis. Some chromosomes fail to recruit Nnf1 and remain unsegregated (arrows). (D) Quantitation of Nnf1 localization. For each genetic background, the percentage of cells harbouring unsegregated chromosomes is plotted; the superimposed colour code specifies the pattern of Nnf1-GFP signal in those cells. See Methods for definitions of ‘stable’ and ‘unstable’. Labels as in Supplementary Fig. 1. (wt-bqt1Δ p = 0.0014).