Binding of ¹²⁵I-Lys-plasminogen to RAW264.7 macrophages is inhibited by anti–annexin II.

(Left panel) Adherent cells (10⁵/well) were washed with DPBS (3 ×). The media were replaced with cold MSFM alone or MSFM containing 0.1 to 6 μg/mL ¹²⁵I-Lys-plasminogen (¹²⁵Lys-Plg). Following incubation for 1 hour at 4°C with¹²⁵I-Lys-plasminogen, cells were washed (3 ×) with DPBS and dissolved in 0.5 NaOH. Data represent the mean ± SE of 3 separate samples. (Right panel) Adherent cells (10⁵/well) were washed with DPBS (3 ×). The media were replaced with cold MSFM alone or MSFM containing monoclonal anti–annexin II IgG (10-25 μg/mL; clone Z014, Zymed), mouse IgG (25 μg/mL), or 25 mM ε-ACA. The cells were incubated with the antibodies for 2 hours at room temperature before the addition of ¹²⁵I-Lys-plasminogen (1.0 μg/mL). Cells were then incubated 1 hour at 4°C. The data represent the mean ± SEM of 6 replicate samples. nIgG indicates normal IgG.