Induction of apoptosis by anti-RLIP76 C-ter antibodies in EAhy926 endothelial cells.(A,C) Flow cytometric analysis after double staining with annexin V/propidium iodide of untreated control cells; cells incubated for 30 minutes, 6 hours, and 24 hours with anti-RLIP76 C-ter antibodies; and in cells treated with H₂O₂ and then incubated additional 30 minutes, 6 hours, and 24 hours in fresh medium or with medium containing anti-RLIP76 C-ter antibodies. (B,D) Cells were also pretreated with α-TCPH. (A,B) Results obtained from 3 independent experiments are reported as mean plus or minus SD. (C,D) Dot plots from a representative experiment. Numbers represent the percentage of annexin V single positive (early apoptosis, bottom right quadrant) or annexin V/PI double positive cells (late apoptosis, bottom right quadrant). Statistical analysis performed by Student t test indicated P < .01 for: control untreated cells versus H₂O₂-treated cells; control untreated cells versus cells incubated for 30 minutes, 6 hours, and 24 hours with anti-RLIP76 C-ter antibodies; and cells treated with H₂O₂ and then incubated in fresh medium versus cells treated with H₂O₂ and then incubated for 6 hours and 24 hours with medium containing anti-RLIP76 C-ter antibodies. P < .01 for any treatment (panel A) versus the same treatment performed after α-TCPH preincubation (panel B).