Fig. 3.

Mitochondrial superoxide and MPO-derived ROS from neutrophils and monocytes are generated on the early onset, whereas NO and NOX-derived ROS from neutrophils, lymphocytes and monocytes are generated on the later in TNFα-activated leukocytes. Human leukocytes from healthy donors (n = 6) were activated by TNFα (10 ng/mL) and incubated for the indicated time periods. (A and B) Total ROS, NO and mitochondrial superoxide were measured as described in methods. Each panel are representative of 6 independent experiments. Results in right panel of each row are plotted as the mean fluorescent intensity of each sample on the vertical axis with mean ± SEM. *p < 0.001 vs. control, ANOVA and Bonferroni post-hoc test. (C and D) MPO and NOX activities from leukocytes (C) or isolated neutrophils, lymphocytes and monocyte (D) were measured as described in methods (n = 6, mean ± SEM). *p < 0.001 vs. control, ANOVA and Bonferroni post-hoc test.