Figure 6

Expression of CD59 and CD105 on annexin V-binding HUVEC microparticles. HUVEC at 48 hrs in culture were incubated for 24 hrs with 5 μM CPT. After incubation MP in culture medium were harvested and double-labeled with annexin V-FITC and a saturating concentration of a PE-conjugated Mab : IgG2a isotype control (IgG-IC, filled area), binding of the isotype control corresponded to the fluorescence of nonlabeled MP; Mab to CD59 (dashed line); Mab to CD105 (thick line). After incubation and washing, samples were analyzed by flow cytometry. The annexin V-FITC positive MP were gated in R1 gate in FITC-flourescence vs. SSC plot (A) and their binding of PE-labeled Mabs was evaluated in PE-fluorescence histogram plot (B). Results are representative of six experiments.