Effect of Plg-RKT on MMP activation. C57BL/6 mice were injected intravenously with 500 μg of either mAb 7H1 or isotype control. After 30 minutes, thioglycollate was injected intraperitoneally. A second injection of Ab was given 24 hours later. After 72 hours, peritoneal lavage fluid from 5 mice/group was collected and pooled. Then, 25 μL of peritoneal lavage fluid from each treatment condition was electrophoresed on 10% SDS gels containing 0.1% gelatin and evaluated as described in “Zymography”: (A) 25 μL was electrophoresed on 4%-20% SDS gels and Western blotted with either anti–MMP-2 (B) or anti–MMP-9 (C). U937 cells (1 × 106 cells/mL) were incubated with plasminogen (200nM) in the presence of either mAb 7H1 (140nM) or isotype control (140nM) overnight at 37°C. Conditioned media (25 μL) was Western blotted with either anti–MMP-2 (D) or anti–MMP-9 (E).
