Figure 4

15d-PGJ2 cytotoxicity involves free radical production and is influenced by intracellular glutathione levels. (A) Time course of 15d-PGJ2-induced ROS production. mOP cells were preloaded with the fluorescent ROS probe DCFDA for 30 min, and then treated with 15d-PGJ2 (10 μM) for 1 h. ROS production was expressed as DCFDA fluorescence intensity (pixel value). (B) mOP cells were pre-treated or not with NAC (1 mM), Ascorbic acid (1 mM) or α-tocopherol (1 mM) for 1 h prior to treatment of 15d-PGJ2 (5 μM) for 24 h, toxicity was examined by counting the apoptotic cells with condensed nuclei. (C) mOP cells were treated or not with BSO (100 μM) for 4 h and the total level of intracellular GSH was measured. (D) mOP cells were treated with BSO (100 μM) for 1 h and then co-treated with 15d-PGJ2 (1 μM) for 24 h. Cells treated with BSO or 15d-PGJ2 alone or untreated were included as controls. Toxicity was examined by counting the apoptotic cells with condensed nuclei. Asterisks indicate significant difference (One-way ANOVA with Newman-Keuls or Dunnet posttest, or two way ANOVA with Bonferroni posttest, *(p < 0.05), **(p < 0.01) ***(p < 0.001); two-tailed t-test used in C).