Figure 7

Activation of NADPH oxidase and ROS production is upstream of NO production in BV-2 microglial cells. Cells were pretreated with (A) DPI (0.1 to 1.0 μM) or (B) AEBSF (6.25 to 200 μM) for 1 h prior to exposure to IFNγ (10 ng/ml) for 12 h, followed by measurement of ROS with CM-H2DCFDA as described in text. Cells were pretreated with (C) DPI (0.05 to 0.2 μM), (D) AEBSF (12.5 to 100 μM), or (E) apocynin (50 to 400 μM) for 1 h prior to exposure to IFNγ (10 ng/ml) for 16 h, followed by measure of nitrite with Griess reaction protocol as described in the text. Results are expressed as the mean ± SEM (n = 3), and significant difference from the respective IFNγ-stimulated group was determined by one-way ANOVA followed by Dunnett’s tests, *P < 0.05; **P < 0.01.