Figure 4.

The Interaction between BUBR1 and Blinkin Is Required for a Functional SAC

(A) Synchronization protocol for RNAi depletion of BUBR1 and rescue with mutants. Time is shown in days (D).

(B) Western blot for BUBR1 showing depletion of endogenous BUBR1 and level of expression of the different mutants (see also Figure S3).

(C) Plot showing time spent in mitosis in the presence of 100 nM nocodazole with each dot representing a single cell analyzed by time-lapse microscopy and bar indicating the median. Time is scored as the time from nuclear envelope breakdown (NEBD) to time of mitotic exit and for each condition 60 cells have been analyzed. The statistical significance is indicated above as determined by the Mann-Whitney test. The time spent in mitosis in the Luciferase control is underestimated as 49 out of 60 cells did not exit mitosis during filming.

(D) Representative still images from the different cell lines analyzed. Time is in minutes and t = 0 is at NEBD.

(E) Synchronization protocol used for the immunodepletion experiment.

(F) Immunopurified BUBR1 complexes analyzed for the presence of Blinkin, Cdc20, Mad2, and Bub3. In the input the asterisks indicates an unspecific band from previous probing of the blot.