Effects of soluble forms of recombinant tumor necrosis factor-related apoptosis-inducing ligand:iron superoxide dismutase (sTRAIL:FeSOD) on intracellular H
, glutathione (GSH), superoxide radical (O
-) and reactive oxygen species (ROS) levels. (A) and (B) After treatment with sTRAIL:FeSOD (1,000 ng/ml), sTRAIL:mFeSOD (1,000 ng/ml) or FeSOD (500 ng/ml) for 3 hours, dihydroethidium (DHE) or dichlorodihydrofluorescein diacetate (DCFDA) was added for detection. (C) through (J) After cells were treated with sTRAIL:FeSOD (1,000 ng/ml) or sTRAIL:mFeSOD (1,000 ng/ml) for 0, 1, 2, 3, 4, 5 or 6 hours, intracellular H2O2, GSH, O2- and ROS were measured by DHR123, NDA, DHE and DCFDA, respectively. The fluorescence was measured by flow cytometry. For each sample, 10,000 events were acquired. Results are expressed as the mean fluorescence intensity. Each bar represents the mean ± SE obtained from three independent experiments (*P < 0.05 vs. untreated control).