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Fig. 3. Bub1 is required for kinetochore localization of BubR1, Cenp-F, Cenp-E and Mad2. DLD-1 or DLD-1 Myc-Mad2 cells were transfected with control or Bub1 siRNA duplexes, treated with nocodazole and then fixed and stained to detect BubR1 (blue), centromeres/kinetochores (ACA, green), DNA and BubR1, Myc-Mad2, Cenp-E or Cenp-F (red). The cells were then analysed by deconvolution microscopy and image stacks projected onto a single plane. (A) Representative projected image stacks showing that repression of Bub1 reduces kinetochore localization of BubR1, Mad2, Cenp-E and Cenp-F. Scale bar, 5 µm. (B) Normalized pixel intensities in control (black bar) and Bub1-repressed (white bar) cells. Values represent the mean and s.e.m. of at least 30 kinetochores in three different cells.





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