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. 2009 Jun 1;185(5):841–858. doi: 10.1083/jcb.200902128

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© 2009 Klebig et al.

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Figure 6. — Expression of cancer-related Bub1 mutants differentially affects checkpoint efficiency and chromosome congression. (A) Immunofluorescence quantification of total cellular Bub1 mutant levels using Bub1 and GFP antisera. (B) Immunofluorescence quantification of Bub1 mutant levels on kinetochores using GFP antisera. (C) Immunofluorescence images of indicated mitotic cells stained with DAPI (DNA), GFP antisera (green), and CREST antisera (red, kinetochores). (D) Mitotic index of Bub1 mutant cells treated for 16 h with nocodazole normalized to Bub1-wt. (E) Ability to complement congression errors in Bub1 mutant cells treated for 1 h with MG132 normalized to Bub1-wt. (F) Quantification of TUNEL-positive cells after nocodazole and Bub1 RNAi. (G) Immunofluorescence quantification of Mad1, Mad2, and BubR1 levels on kinetochores in indicated cell lines treated with Bub1 RNAi. Error bars represent standard deviation. (H–J) Immunofluorescence images of the indicated control or Bub1 RNAi–treated prometaphase cells stained with Mad1 (H), Mad2 (I), or BubR1 (J) antisera (green) and CENP-A or CREST antisera (red, kinetochores). Insets show a higher magnification view of a single kinetochore. Bars,10 µm.