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. 2002 Aug 5;158(3):487–496. doi: 10.1083/jcb.200204048

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Copyright © 2002, The Rockefeller University Press

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Figure 4. — BubR1 is required for other spindle checkpoint proteins to associate with chromosomes. (A) Immunoblot analysis of various checkpoint proteins associated with chromosomes. Mitotic chromosomes were isolated from mock-depleted (lanes 1, 2, 5, and 6) or BubR1-depleted (lanes 3, 4, 7, and 8) extracts that were untreated (lanes 1, 3, 5, and 7) or treated with nocodazole (lanes 2, 4, 6, and 8). The extracts or chromosomal fractions were immunoblotted for proteins indicated on the left. The migration of molecular size standards is indicated on the right for BubR1 and Bub1 blots. (B) Phosphatase treatment removes mobility shift of chromosomal BubR1 and Bub1. Chromosomal pellets prepared from untreated extracts (lanes 1, 3, 5, and 7) or extracts treated with nocodazole (lanes 2, 4, 6, and 8) were treated with phosphatase buffer (lanes 1, 2, 5, and 6) or LPP (lanes 3, 4, 7, and 8). The samples were then immunoblotted for BubR1 (lanes 1–4) or Bub1 (lanes 5–8). The migration of molecular size standards is indicated on the right.