Fig. 6. Effects of CD222-derived peptides on mini-Plg-induced apoptosis in ECs. (A) HUVECs were incubated in medium containing 1% serum for 24 hours with mini-Plg (240 nM) in the presence of the CD222-derived peptides and control peptides (20 µg ml^-1), then stained with annexin-V/FITC and analysed under a fluorescence microscope. Apoptotic cells are indicated by arrows. Scale bar, 100 µm. (B) Cells were treated as in A. After incubation, the detached cells were pooled with the trypsinized adherent cells and assayed for the occurrence of apoptosis by FACS analysis using an apoptosis detection kit (Calbiochem, La Jolla, CA, USA). Viable cells were annexin-V negative and PI negative; nonviable necrotic cells or late apoptotic were annexin-V positive and PI positive; early apoptotic cells were annexin-V positive and PI negative. Similar results were obtained in three different experiments.