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. Author manuscript; available in PMC: 2009 Jan 31.
Published in final edited form as: Mol Cell. 2008 May 23;30(4):403–414. doi: 10.1016/j.molcel.2008.03.009

Figure 1. Analysis of Fbw8/IRS-1 Interaction In Vivo.

Figure 1

(A and B) Analysis of the Flag-Fbw8 immunoprecipitates. Silver stain (A) and immunoblot (B) analysis of Flag-Fbw8-associated proteins, isolated as described in the Supplemental Data.

(C) Interactions between ectopically expressed Fbw8 and IRS-1. HEK293 cells were transfected with vectors expressing V5-IRS-1 and Myc-Fbw8. At 48 hr posttransfection, cells were harvested. Extracts (500 µg) were used for immunoprecipitation with anti-Myc or anti-V5 antibodies followed by separation through 6% SDS-PAGE and immunoblot analysis as indicated.

(D) Association between endogenous IRS-1 and CUL7. Extracts (1 mg of protein) from MCF-7 cells were subject to immunoprecipitation with IgG (lane 1) or anti-CUL7 (Mab38) antibodies (lane 2). The precipitates were separated by 6% SDS-PAGE followed by immunoblot analysis.