Decreased cell viability and ROS production induced by oxidative treatment.

<p>(A) Twenty-four hours after exposure to diverse concentrations of H₂O₂ for 30 min, the viability of WRL68 cells was analysed with the MTT assay. Cell viability was expressed as the percentage of viable cells with respect to those determined in the controls (H₂O₂ non-treated cells), which was considered to be 100%. Bar graph shows the mean ± SEM of three independent experiments. Significant differences with respect to the controls (H₂O₂ non-treated cells): *<i>P</i> < 0.05, **<i>P</i> < 0.01 and ***<i>P</i> < 0.001. (B) ROS production by WRL68 cells was evaluated by incubating the cells with H₂DCFDA for 30 min, before exposing them to H₂O₂. The level of DCF fluorescence was evaluated by flow cytometry at 0, 5, 15 and 30 min during exposure and at 15 and 30 min after 30 min exposure to H₂O₂ (time point of 45 min and 60 min, respectively, in the figure). The linear graph shows the percentage of fluorescent cells (DCF positive cells) at each time point. Mean ± SEM of three independent experiments. Significant differences between each treatment and their respective controls (H₂O₂ non-treated cells): *<i>P</i> < 0.05, **<i>P</i> < 0.01, and ***<i>P</i> < 0.001. (C) Bar graph showing the proportion of DCF-fluorescent cells (indicating ROS production) at 15 minutes of exposure to different doses of H₂O₂ treatment. Bars represent the mean ± SEM of three independent experiments. Significant differences with respect to H₂O₂ non-treated cells: ***<i>P</i> < 0.001.