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. 2007 Aug 6;204(8):1825–1835. doi: 10.1084/jem.20070872

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Copyright © 2007, The Rockefeller University Press

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Figure 1. — Physical and functional association between FBW7 and NOTCH1. (A) Schematic depiction of the NOTCH1 (N1) protein domains, and N1-IC WT (N1-IC^WT) and 82 and 162 ΔPEST mutants (N1-IC^Δ82 and N1-IC^Δ162). Asterisk denotes L1601P mutation. The FBW7 degron is highlighted in gray. LNR, LIN-Notch repeat domain; RAM, RBJ-κ–associated module; TAD, transcriptional activation domain; PEST, P-E-S-T–rich domain; HD, heterodimerization domain. (B and C) Coimmunoprecipitation experiments using anti-Flag beads or anti-HA beads, showing interaction between DN FBW7 and N1-IC^WT, but DN FBW7 (D) does not interact with N1-IC^Δ82 and N1-IC^Δ162. (E) Cotransfection with FBW7α repressed N1^L1601P (P < 0.02), but not N1^L1601P-Δ82, in reporter assays using a CSL luciferase reporter. pCDNA3.1 represents the control, “empty” expression vector. Error bars represent the SD of duplicate wells.