FIGURE 2:

AGP enhances ROS accumulation in melanoma cells. (a) AGP enhances ROS levels in melanoma cells (Mel-RM, Mel007) but not normal cells (melanocytes, human fetal lung fibroblasts). Cells were treated with AGP (5, 15, 30 s). ROS level was quantified by the fluorescent dye CM-H2DCFDA and is shown as a fold change relative to control cells treated only with He gas flow. Data were normalized to ROS levels in control melanocyte cells. (b) Mel007 or melanocyte cells were treated with AGP (5, 15, 30s) or pretreated with NAC (1–2 h) followed by AGP treatment (5, 15, 30 s). ROS level was determined as in a. (c) Mel007 or melanocyte cells were treated with AGP or pretreated with NAC (1–2 h), followed by AGP treatment. GSH level was quantified as described in Materials and Methods. (d) Mel007 or melanocyte cells were treated with AGP (5, 15, 30 s) or pretreated with NAC (1–2 h), followed by AGP treatment (5, 15, 30 s). Nitric oxide was quantified after cells were labeled with DAF-FM diacetate. All values are mean ± SD of three independent experiments performed in triplicate. *p ≤ 0.01, **p ≤ 0.001; ANOVA.