Binding of anti-IIbβ3 MoAb and soluble fibrinogen to TPA-induced CMK cells. D3-adherent, D3-detached, and control CMK cells were incubated with (A) anti-IIbβ3 complex MoAb or (B) 200 μg/mL FXS-labeled fibrinogen for 30 minutes at room temperature, and flow cytometry was performed. D3-adherent cells were harvested by detaching them mechanically after the binding reaction. (C) D3-adherent and D3-detached cells were pretreated with 40 μg/mL of anti-IIbβ3 MoAb or control mouse IgG for 5 minutes at room temperature and then treated with 50 μg/mL FXS-labeled fibrinogen, after which flow cytometry was performed. The MFI of FXS-labeled fibrinogen bound to D3-adherent cells was taken as 100%, and results represent the mean ± SD of 3 independent experiments. (D) D3-adherent, D3-detached, and control CMK cells were incubated with the activation-dependent anti-IIbβ3 MoAb, PAC1 as described above, and flow cytometry was performed. Results of (A), (B), and (D) are representative of 3 experiments.