Figure 4.

Effects of NCTD on phosphorylation of integrin β₃, Src and FAK on a fibrinogen‐coated surface. Washed human platelets were preincubated with NCTD (0.15 and 0.3 μM) or the solvent control (0.1% DMSO) and subsequently activated by immobilized fibrinogen (100 μg/ml). Platelets were collected, and their subcellular extracts were analysed to determine the levels of (A) integrin β₃, (C) Src and (D) FAK phosphorylation. (B) For immunoprecipitation study, washed platelets were preincubated with 0.3 μM NCTD or the solvent control (0.1% DMSO) for 3 min. and then allowed to spread on immobilized fibrinogen (100 μg/ml). Next, the platelets were lysed, Protein G Mag Sepharose Xtra beads (10 μl) were added with the anti‐integrin β₃ mAb (1 μg/ml), and the platelets were incubated overnight with rotation for immunoblotting. The profiles in (A) and (B) represent four independent experiments; all data are presented as means ± standard errors of the means (n = 4). **P < 0.01, compared with the immobilized bovine serum albumin (BSA)‐treated group; ^# P < 0.05 and ^## P < 0.01, compared with the immobilized fibrinogen‐treated group.