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. 2010 Jul 12;190(1):89–100. doi: 10.1083/jcb.201001050

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© 2010 Maciejowski et al.

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Figure 3. — Mps1 directs the assembly of Cdc20 inhibitory complexes in interphase. (A) Mps1^wt and Mps1^as cells were treated with 3MB-PP1 for 2 h. Extracts were immunoprecipitated with antibodies to Cdc20 and resolved by SDS-PAGE. Levels of BubR1, APC8, Mad2, and Cdc20 in each IP were determined by quantitative immunoblotting. The ratio of each protein to Cdc20 was computed and normalized to ratios obtained from untreated Mps1^wt cells (= 1.0). (B) Mps1^wt cells were treated with ZM for 2 h and processed as in A. White line indicates that intervening lanes have been spliced out. (C) Interphase and mitotic Cdc20 IPs were electrophoresed on 6% gels to resolve mitotically phosphorylated APC8 (left) and BubR1 (right) polypeptides as distinct species. Numbers at the bottom are loading amounts. Black lines indicate that intervening lanes have been spliced out. ctrl, control. Error bars indicate SEM.