Soluble fibrinogen binding to _Vβ₃ and _Vβ₃(T562N) transfected cells. Cells were preincubated with 1 mmol/L RGDW, 1 mmol/L manganese, or buffer with 10 μg/mL of anti-_V antibody, LM142. After 30 minutes of incubation, cells were washed and then incubated with 150 μg/mL FITC-conjugated fibrinogen and PE-conjugated antimouse IgG for 30 minutes and analyzed by flow cytometry. (A) Dot blots represent FITC-fibrinogen (horizontal) and LM142 (vertical) binding in the absence of RGDW and manganese. (B) Fibrinogen binding to cells expressing high levels of _Vβ₃ (denoted by the rectangle in the dot blots) was analyzed on the histograms. (a through c) Wild-type _Vβ₃ transfected cells; (d through f) _Vβ₃(T562N) transfected cells. (a and d) With RGDW; (b and e) with buffer; (c and f) with manganese.