Anti–annexin II inhibits plasmin binding to human monocyte-derived macrophages.
Cells were aliquoted into 96-well plates (105/well). The next day 2 groups of cells were washed (3 ×) with DPBS. The media were replaced with MSFM containing monoclonal anti–annexin II (clone Z014, Zymed) or normal mouse IgG (nIgG; 25 μg/mL). Following incubation for 30 minutes at 37°C, plasmin (1 μg/mL) was added and cells were incubated for 1 hour at 4°C. The remaining groups of cells were washed and media replaced with plasmin (Pls) alone, plasmin and ε-ACA (25 mM), or plasmin and purified annexin II. Following incubation for 1 hour at 4°C, membrane-bound plasmin was determined as described in “Materials and methods.” The data represent the mean ± SEM of 3 replicate samples. Control (Ctrl) indicates cells in the absence of exogenous plasmin.
