Fig. 1.
Effects of RvE1 on neutrophil apoptosis. Human neutrophils (5 × 106 cells/mL) were cultured for 24 h with RvE1 and viability, mitochondrial transmembrane potential (ΔΨm) [chloromethyl-X-rosamine (CMXRos) staining] and apoptosis (annexin-V–FITC binding and nuclear DNA content) were assessed. (A) RVE1 at high concentrations suppresses intrinsic apoptosis. Data are means ± SEM (n = 4–11). *P < 0.05 vs. untreated. (B) To monitor ROS production, neutrophils were loaded with H2DCFDA (5 μM) and then left untreated (C, control) or challenged with RvE1 for 15 min. Data are means ± SEM (n = 4–7). *P < 0.05; **P < 0.01; ***P < 0.001 vs. untreated. (C) Caspase-8 activity was assessed at 4 h post-RvE1 with flow cytometry using FITC-labeled z-Ile-Glu(Ome)-Thr-Asp(Ome)-fluoromethylketone (z-IETD-fmk) as a substrate. The effect of the Fas-activating antibody (CH-11 Ab) is shown for comparison. (n = 4–7). *P < 0.05; **P < 0.01 vs. untreated.