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Figure 5


Fig. 5. Confirmation of INCENP as a P190-reactive phosphoprotein. (A) INCENP and Aurora-B are specifically immunoprecipitated from HeLa mitotic cell lysates using the P190 antibody. (B) P190 recognizes bands corresponding to the molecular weight of INCENP (arrow) in both INCENP and Aurora-B IPs. (C) Site-directed mutagenesis of serine 197 to alanine (S197A) or aspartate (S197D) prevents P190 binding. The arrow shows the location of GFP-INCENP. (D) Mutation of serine 197 to alanine, but not to aspartate, prevents INCENP from properly localizing to the midbody. The arrow denotes the midbody in the merged panel and the region containing the midbody was enlarged to clearly demonstrate that S197A (green) is not present in the midbody containing Aurora-B (red). Bars, 10 µm.





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