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. 2011 Aug 2;286(38):33613–33621. doi: 10.1074/jbc.M111.245126

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© 2011 by The American Society for Biochemistry and Molecular Biology, Inc.

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FIGURE 5. — Ala-153 mutations of KLHL7 inhibit Cul3 interaction but not dimerization. A, HCT116 cells were transfected with either FLAG-KLHL7 wild-type or missense mutants. The cell lysates were immunoprecipitated (IP) with anti-FLAG antibody and immunoblotted with anti-Cul3 and anti-FLAG antibodies. B, HeLa cells cultured on coverslips were transfected with HA-Cul3 and FLAG-KLHL7 A153V. The cells were fixed and stained with anti-HA and anti-FLAG antibodies and Hoechst 33342. C, FLAG-KLHL7 (wild-type (WT) and A153V (mt)) and HA-KLHL7 (wild-type (WT) and A153V (mt)) were expressed in HCT116 cells as indicated. FLAG-KLHL7 was immunoprecipitated with anti-FLAG antibody and immunoblotted with anti-HA and anti-FLAG antibodies. D, HeLa cells on coverslips were transfected with FLAG-KLHL7 and HA-KLHL7 A153V. After fixation, cells were stained with anti-FLAG and anti-HA antibodies and Hoechst 33342. Scale bars, 20 μm.