Figure 3.
Activated thrombin-activatable fibrinolysis inhibitor (TAFIa)-associated propagation of plasminogen accumulation initiated from the activated platelet-surfaces. (A) The left panel is a 15-minute “control” panel. Four concentric circles in the dense fibrin network region were analyzed based on Alexa Fluor 488-labeled fibrinogen fluorescence; green (right). Concentric circles have diameters of 15, 30, 45, and 60 pixels (17, 34, 51, and 68 µm, respectively). (B) Fluorescence intensities (FI) of Alexa Fluor 568-labeled plasminogen (plg-568) were measured in 4 concentric circles in the dense fibrin network region, and the average FI for the center and the 3 donut-shaped areas were determined. Representative FI changes for plg-568 are shown relative to the maximum FI at the center (RFI %). The shading of the lines is from the center (ie, activated platelets to the outer [light red line to gray lines]). Control (a), TAFIa inhibitor (TAFIaI) (b), neutralizing antibody for thrombomodulin (TM-Ab) (c-d). (C) Propagation of plasminogen accumulation is represented as the time delay in peak plasminogen FI from the center to each outer donut-shaped area (definition is indicated in the dotted square areas, as D1, D2, and D3). D1, D2, and D3 are presented as the median and interquartile range. Control (n = 23) (a), TAFIaI (n = 21) (b), TM-Ab (n = 17) (c). Significance was analyzed by the Mann-Whitney U test (*P < .01 vs control, **P < .01 vs TAFIaI).