cVWF release from microthrombi. Microthrombi were exposed to plasmin (75 μg/mL) or buffer. (A-C) Platelet-free supernatants were collected and analyzed by ELISA. Next, preformed microthrombi were separated from soluble VWF by centrifugation and subjected to either degradation by plasmin (75 μg/mL) or buffer. Platelet complexes were removed by a second centrifugation step and supernatants analyzed by western blotting (D) and ELISA (E-G). Data represent 3 independently executed experiments. Bar graphs show means ± SD. No Plt, no platelets; Plm, plasmin.