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. 2010 Sep 6;190(5):835–852. doi: 10.1083/jcb.201002070

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© 2010 Petrovic et al.

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Figure 3. — Interaction of MIS12C with NDC80C. (A) Schematic representation of the NDC80C. (B) Binding curves were measured by ITC by titrating NDC80C (left) or SPC24^57–197–SPC25^70–224 (right) in a cell containing MIS12C^NSL1-258. (C) Size-exclusion chromatography elution profiles and SDS-PAGE analysis of recombinant NDC80C expressed in, and purified from, insect cells (top), MIS12C^NSL1-258 (middle), and their stoichiometric combination (bottom). (D) Elution profiles and SDS-PAGE analysis of NDC80C (top), MIS12C^NSL1-227 (middle), and their stoichiometric combination (bottom). The MIS12C^NSL1-227 construct binds NDC80C. (E) As in C, but with the kinetochore-binding portion of NDC80C, the SPC24^57–197–SPC25^70–224 construct. (C–E) Dashed gray lines and numbers indicate elution markers in the size-exclusion chromatography experiments and their molecular masses (in kilodaltons), respectively.