Figure 3.

Bub1 is indispensable for localization of Mad2 on cohesion-defective kinetochores. The cut9 single mutant, cut9 mis4 double mutant, and cut9 mis4 Δbub1 triple mutant cells were incubated at the restrictive temperature for two generations (5, 6, and 8 h, respectively), and fixed for immunostaining of Cnp1 (s.p. CENP-A). Mad2 was visualized by GFP fusion in these cells. Stacks of Z-sections were taken and flattened after deconvolution. In merge panels, Mad2, Cnp1, and DNA are colored green, red, and blue, respectively. Bar, 5 μm.