Identification of activators and cellular activation of plasminogen. (A) Fibrin autography of platelets, cortical neurons, HMEC-1 cells, and THP-1 cells. The samples were electrophoresed on 8% (wt/vol) polyacrylamide, SDS was then exchanged with 2.5% (wt/vol) Triton X-100, and the gel was overlaid on a fibrin-agarose indicator gel. The picture was taken after 4 hours at 37°C. The position of purified controls (Pn indicates plasmin; tPA, and uPA) is indicated on top. The thin vertical line indicates assembly from the same gel. The thick vertical line separates 2 different gels. (B) THP-1 cells (105 cells/well) were incubated with various concentrations of plasminogen (0-3μM) and 0.75mM CBS0065. Kinetics of plasmin formation (mOD/minute) was followed by measuring the release of p-nitroaniline. Data were fitted according to the Michaelis-Menten equation (Km = 492nM).
