FIGURE 8:

Bub1 removal from sister centromeres after capture. Cells expressing Bub1-3×mCherry, GFP-Tub1, and TetR-GFP and bearing MET3pr-CDC20 and GAL1pr-CEN3-TetOs were treated as described in Figure 7. Representative deconvolved, Gaussian-filtered MIPs are shown in A. Scale bars, 1 μm. (A) Time series showing Bub1 disappearance from sister centromeres after capture. The blue dashed boxes surrounding the first frame highlights the corresponding regions of the expanded views shown in Supplemental Figure S7D and Supplemental Video S10. (B) Kymograph from the cell in A, made from 15 by 15 boxes approximately centered on the centromeres (top). (C) Bub1 intensity at the centromeres in A and B normalized by taking the ratio of the mCherry signal to the associated GFP intensity (dots). A color-coded sliding window average of five frames is also shown. (D) Timing of the final disappearance of Bub1-3×mCherry and Mad1-3×mCherry from centromeres relative to the completion of capture. The completion of capture was defined as the first frame in which the centromeres could no longer be resolved from the spindle. Error bars represent SD. *p < 0.05.