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Fig. 7. Aurora B is required for localization of Bub1. DLD-1 cells were transfected with control or Aurora B siRNA duplexes, treated with nocodazole and then fixed and stained to detect Aurora B (blue), centromeres/kinetochores (ACA, green) DNA and Bub1, BubR1 or Cenp-E (red). (A) Projection of a deconvolved image stack showing repressed and unrepressed prometaphase DLD-1 cells transfected with Aurora B siRNA duplexes in the same field of view. (B) Normalized pixel intensities in nocodazole-treated control (black bar) and Aurora-B-repressed (white bar) cells. Values represent the mean and s.e.m. of at least 30 kinetochores in three different cells. (C) Repression of Aurora B inhibits the localization of Bub1 during prophase. Projections of deconvolved image stacks showing prophase DLD-1 cells transfected with control (scramble) and Aurora B siRNA duplexes, fixed and stained, as described above. (D) Standard immunofluorescence of prometaphase DLD-1 cells that were transfected with control, Bub1, BubR1 or Cenp-E siRNA duplexes, fixed and stained to detect DNA, Aurora B (red) and Bub1, BubR1 and Cenp-E (Green). Representative examples showing that repression of either Bub1, BubR1 or Cenp-E has no apparent effect on Aurora B localization.





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