Depletion of S100A10 results in decreased endothelial cell plasminogen binding and plasmin generation. To detect the total cellular levels of annexin A2 and S100A10, primary murine endothelial cells, isolated from WT or S100A10^−/− mice (A), as well as control and S100A10 depleted TIME cells (B), were dissociated from culture flasks, lysed, subjected to SDS-PAGE, and immunoblotted with antiactin (loading control), antiannexin A2, or anti-S100A10 antibodies. Cell-surface protein levels for primary murine endothelial cells (C) and TIME cells (D), as detected by cell-surface biotinylation, are shown. FITC-Pg binding to the primary murine endothelial cells (E) or TIME cells (F) was measured by FACS. Quantification of flow cytometric analysis of Pg binding was calculated with WinMDI software. Loss of S100A10 affected tPA-dependent plasmin generation by primary murine endothelial cells (G) and TIME cells (H) and uPA-dependent plasmin generation by TIME cells (I). Statistical analysis was performed with the Student t test (E,G) or ANOVA (F,H); *P < .1, **P < .01, and ***P < .001. Figures were generated with Adobe Photoshop CS3 Version 10 (Adobe Systems Incorporated).