Identification of plasminogen binding proteins in RAW264.7 macrophage membrane preparation.

For ligand blot, cell lysates and isolated membranes (Mem) were electrophoresed in gradient polyacrylamide gels and transferred to a PVDF membrane. PVDF membranes were blocked and subsequently incubated with buffer alone (Ctrl) or buffer containing 10 μg/mL Lys-plasminogen (+Lys-Plg). Bound plasminogen was visualized using monoclonal antiplasminogen IgG as described in “Materials and methods.” The presence of immunoreactive annexin II in macrophage lysates and isolated membranes was determined by Western blot as described in “Materials and methods.”