Apoptotic pathway induced by anti-RLIP76 C-ter in EAhy926 cell line. (A) Flow cytometric data were obtained in the absence (first row) or in the presence (second row) of 30 μM pan-caspase inhibitor zVAD. The numbers in each panel refer to the percentage of cells containing caspase 3 in its active form. Results obtained in a representative experiment are reported. (B,C) Graphs showing the mean plus or minus SD of the percentages of cells with the active form of caspase 3 obtained from 3 different experiments done without (panel B) or with (panel C) zVAD. Statistical analyses indicate a significant (P < .01) decrease in caspase 3 activity in cells pretreated with zVAD before anti-RLIP76 C-ter antibody exposure. (D) Quantitative flow cytometric analysis investigating the JNK activation state 6 hours after the various treatments with a polyclonal antibody able to identify JNK (pT183/pY185). The numbers in each panel refer to the percentage of cells containing JNK in its active form. Results obtained in a representative experiment are reported. (E) Time-course analysis of the activation state of JNK. Statistical analyses of the results obtained from 3 independent experiments (reported as mean ± SD) indicate a significant difference (P < .01) between cells treated with RLIP76 C-ter antibodies, alone or in combination with H₂O₂, and both untreated and H₂O₂-treated cells at any time point considered.