Figure 6
NADPH oxidase inhibitors and an inhibitor of de novo protein synthesis, cyclohexamide, attenuate ROS, but only at later time points after initiation of the MPP+ treatment. (A) N27 cells were treated with 300 μM MPP+ in the absence or presence of 10 nM PAO. Intracellular ROS levels were measured using carboxy-H2-DCFDA and flow cytometry at 3, 6, or 24 hours after initiation of the MPP+ treatment. * represents p < 0.001 compared to 3 hours MPP+ and # represents p < 0.001 compared to 24 hours MPP+. (B) N27 cells were treated with MPP+ for 3 or 6 hours in the absence or presence of the protein synthesis inhibitor cyclohexamide (c-hex). Intracellular H2O2 levels were measured as described above. Treatments are plotted as a percent of the 3-hour MPP+ treatment. * represents p < 0.001 compared to 3 hours MPP+ and # represents p < 0.05 compared to 6 hours MPP+. Data are from 3 independent experiments with n = 6 wells per experiment.