Expression of DAF, CD59, and MCP on HUVECs and DMECs. (A) Expression of DAF, CD59, and MCP on resting ECs was assessed by flow cytometry using mAbs 5B2, A35, and TRA-2-10, respectively. The figure shows expression on DMECs and HUVECs with background fluorescence (FITC-labeled rabbit anti–mouse Ab alone, …) and specific antigen expression (—). (B) EC monolayers were incubated for 30 minutes in the absence or presence of PIPLC before harvesting and analysis by flow cytometry. DAF and CD59 were detected with MoAb 5B2 and A35, respectively. The figure shows background fluorescence (…), untreated ECs demonstrating constitutive expression (shaded histogram), and PIPLC-treated ECs (—). (C) HUVECs were harvested with either trypsin/EDTA (▪) or a nonenzymatic cell-dissociation solution (▨) before analysis by flow cytometry. DAF, CD59, and MCP were detected with MoAbs 5B2, A35, and TRA-2-10, respectively. The figure is representative of 3 similar experiments.