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Figure 5 | Cell Communication and Signaling

Figure 5

From: Rac1 activation inhibits E-cadherin-mediated adherens junctions via binding to IQGAP1 in pancreatic carcinoma cells

Figure 5

Interaction of IQGAP1 with Rac1 or β-catenin. (A) IQGAP1 was precipitated from lysates of EGFP-, EGFP-Rac1(V12)- and EGFP-Rac1(N17)-expressing PANC-1 cells and coprecipitated β-catenin was determined by Western blotting (left panel). In parallel, immunoprecipitates of EGFP/EGFP-Rac1 proteins were analysed for coprecipitated IQGAP1 (right panel). Staining of the precipitated proteins verified equal amounts of protein in each immunoprecipitation (lower panels) (B) Binding of IQGAP1 to β-catenin was confirmed by GST-pull down assays. GST-tagged β-catenin was incubated with 500 μg of lysate prepared from EGFP-, EGFP-Rac1(V12)- or EGFP-Rac1(N17)-expressing PANC-1 cells and GST-β-catenin complexes were precipitated by binding to glutathione sepharose. Precipitated IQGAP1 and GST-β-catenin was determined by Western blotting. Determination of IQGAP1 and EGFP proteins in 30 μg of the cell lysate served to demonstrate equal amounts of protein in each sample. One representative blot of three independent experiments is shown.

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