Effects of glycemia on plasmin activity. (A) Time course of incubation of plasma-purified plasminogen with tPA in the absence of fibrin was performed to ensure complete conversion to plasmin. Samples were analyzed by western blot using an anti-plasminogen antibody for detection. (B) S2251 chromogenic substrate (800 μm/L) analysis of plasmin protease activity using plasmin generated by preincubation with tPA of control plasminogen (n = 9), protein purified from T1DM plasma at baseline (pre, n = 9), or after improving glycemic control (post, n = 9). (C) Analysis of plasmin protease kinetics with K_cat/K_m values using substrate concentrations between 50 and 1600 μmol/L. PLM, plasmin; PLG, plasminogen.