The 14-3-3ζ–c-Src–integrin-β3 complex in platelets. (A) Washed human platelets were plated on fibrinogen-coated coverslips at 37°C for 20 minutes. Confocal microscopy of permeabilized platelets stained with antibodies against integrin-β3 (blue), 14-3-3ζ (green), and c-Src (red). Arrowheads indicate 14-3-3ζ⁺, c-Src⁺, and integrin-β3⁺ deposits. Scale bar, 10 μm. (B) Pull-down analysis of the interaction between 14-3-3ζ or the 14-3-3ζ mutant and WT c-Src. (C) Pull-down analysis of the interaction between c-Src or the c-Src mutant and WT 14-3-3ζ. (D) Western blot analysis of recombinant c-Src or 14-3-3ζ protein precipitated by biotin-conjugated WT β3CT, β3CT lacking amino acids 759 to 761 (β3CTΔ759) or β3CT lacking amino acids 749 to 753 (β3CTΔEATST). (E) Western blot analysis of recombinant c-Src or 14-3-3ζ mutant proteins precipitated by biotin-conjugated β3CT. Δ1, Δ2, and Δ3 refer to recombinant 14-3-3ζ proteins lacking amino acids 45 to 58, 112 to 128, and 165 to 180, respectively. Data are shown as means ± SD (n = 3). Statistical significance was determined by Student t test. *P < .05, **P < .01.