FIG 5.

p66^Shc cooperates with FAK to recruit Rho GEFs. (A) HUVECs were exposed to microbeads bound to either BSA or fibronectin (Fn), and a magnetic field was applied for 5 min. The recruitment of p115-RhoGEF and GEF-H1 to adhesion complexes is shown. (B to E) Cells were transduced with control shRNA or shRNA directed against FAK (B, C) or p66^Shc (D, E). Magnetic force was applied to fibronectin-bound microbeads for 5 min, and adhesion complexes were isolated to assess recruitment of p115-RhoGEF (B, D) or GEF-H1 (C, E). (F) Solid-phase His fusions of full-length p66^Shc, p52^Shc, or p66^Shc from which PTB was deleted were used to pull down cell lysates. The captured proteins are indicated. (G) His-p66^Shc was used to pull down lysates in cells transduced with control or FAK shRNA. The results in panels A to G are representative of those from 2 to 4 separate experiments. The numbers to the left of the gels are molecular masses (in kilodaltons).