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. 2006 Dec 4;175(5):743–753. doi: 10.1083/jcb.200605081

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Copyright © 2006, The Rockefeller University Press

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Figure 2. — Confirmation of the binding specificity of the FHA binding proteins of Rad53. (A) Protein extracts from MMS-treated (0.1% MMS for 3 h) Rad9-HA or Mrc1-TAP cells were divided into equal fractions and subjected to pull-down assays using different GST fusion FHA domains as indicated. The GST-FHA domain used in the pull-down assay was stained by Ponceau as a control. Protein extracts from untreated Shs1-TAP, Cdc11-TAP, or Cdc10-HA cells were subjected to the same FHA domain pull-down assays. (B) Binding of the FHA1 domain to septins is Cdc10 dependent. Single-deletion strains for different septins and septin-associated proteins were analyzed for the ability of the FHA1 domain to specifically bind TAP-tagged Cdc11.