FIG 3.

FAK recruits Shc to adhesion complexes. (A) (Left) Microbeads coupled to anti-transferrin receptor antibodies, BSA, or fibronectin were allowed to settle on cells for 40 min, and a magnetic field was applied for the indicated times. Adhesion complexes and lysates were immunoblotted for FAK and FAK (pY397). (Right) The bar graphs show the means ± SEMs from 6 (for FAK) or 5 [for FAK (pY397)] determinations. P was <0.05 at 5 min (FAK) and 10 min [FAK and FAK (pY397)]. (B) Cells were transduced with control or two different FAK shRNAs [FAK and FAK(2)], and fibronectin-coated microbeads were subjected to magnetic tension for 5 min and assessed for recruitment of Shc to adhesion complexes. FAK was knocked down ∼92% [FAK] and ∼70% [FAK(2)]. (C) Cells were transduced with control or Shc shRNA and treated as described in the legend to panel B to assess the recruitment of FAK to adhesion complexes. Shc was knocked down ∼93%. (D) Cells were transduced with either the empty Flag construct or Flag-PTB and assessed for FAK recruitment to adhesion complexes. The results in panels B to D are representative of those from 2 to 4 experiments. The numbers to the left of the gels are molecular masses (in kilodaltons).