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. 2006 Feb 27;172(5):759–769. doi: 10.1083/jcb.200509096

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Copyright © 2006, The Rockefeller University Press

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Figure 4. — The Rac1–β-Pix interaction mediates Rac1 targeting to the plasma membrane. (A) The nongeranylgeranylated V12Rac1C189S mutant is targeted to the plasma membrane in β-Pix–expressing cells. HEK293 cells were transfected with GFPV12Rac1C189S (green) with or without myc–β-Pix (red) and were seeded on fibronectin for 1 h. Cotransfection of β-Pix results in the detection of GFPV12Rac1C189S in membrane ruffles, which is blocked by the Rac1 COOH-terminal peptide (bottom three panels). Bars, 10 μm. (B) Rac1, β-Pix, and RhoGDI can form a trimeric complex. Pull-down experiments were performed with GST or GST–β-Pix in COS-7 cell lysates in the presence of control (Ctrl) or the COOH-terminal Rac1 peptide (Rac1 C-term). RhoGDI and GST–β-Pix were detected by immunoblotting. TCL, total cell lysate.