Cohesin decay destabilizes kinetochore–microtubule interactions. (A) EGFP-BubR1 localization on chromosomes in strain D + TEV (top), strain D + TEV buffer (bottom left, negative control), and strain containing 100% Rad21^TEV + TEV (bottom right, positive control). Times are relative to TEV injection. (B) Relative integrated intensity of EGFP-BubR1 fluorescence in the indicated strains normalized to time of TEV injection (t₁₀/t₀). n = 40/4; n = 56/6; and n = 39/4 (n, number of metaphases analyzed/number of independent embryos). (C) Profile of EGFP-BubR1 levels from a single metaphase of strain D + TEV across time. (D) Mad2-EGFP localization in strain D + TEV (top), D + TEV buffer (bottom left, negative control), and strain containing 100% Rad21^TEV + TEV (bottom right, positive control). Times are relative to TEV injection. (E) Frequency distribution of metaphase plates presenting different numbers of Mad2-EGFP–positive signals per metaphase 10 min after TEV protease or TEV buffer injection. n = 4, 8, and 5 embryos (>100 metaphases were analyzed per condition). (F) Profile of Mad2-EGFP levels from a single metaphase of strain D + TEV across time. For all measurements, only embryos that did not display chromatid disjunction within the course of the experiment (20 min) were analyzed. Mean ± SD. Bars, 5 µm. See also Fig. S2.