Source and kinetics of TNCB induced ROS production <i>in vitro.</i>

<p>(A) DCF fluorescence after incubation of BMDC with different concentrations of TNCB for 2 h either with or without NAC treatment was analyzed by flow cytometry. Data show mean fluorescence intensity of DCF +/− SD of triplicate stimulations. One of two independent experiments is shown. (B) Pam212 or L929 cells were left untreated (−) or stimulated with TNCB (300 µM) after pre-treatment with antioxidants NAC or RF-40s for 1 h. In addition, the influence of the mitochondria specific antioxidant APDC and the NADPH oxidase specific antioxidant DPI was analyzed under the same conditions. DCF fluorescence was analyzed 5, 10, 15, 20, 25, 30, 35, 40 and 45 min after TNCB addition. Data show mean fluorescence as calculated comprising all timepoints (B) or kinetics of fluorescence (C) +/− SD of quadruplicate wells from one representative experiment out of three. (D) gp91^phox −/− mice were sensitized with TNCB (3%) or mock treated with acetone. 5 days later, mice were challenged with TNCB (1%) and increase in ear thickness was measured 24 h later. Data shows mean increase in ear thickness +/− SD of n = 3 mice/group. One representative of three independent experiments is shown.