Research Article
Role of Protein Kinase C and Nox2-Derived Reactive Oxygen Species Formation in the Activation and Maturation of Dendritic Cells by Phorbol Ester and Lipopolysaccharide
Figure 2
Treatment of BM-DCs (2 × 105 cells/well) with PDBu (10 μM) and analysis of the formation of intra- and extracellular ROS measured by (a) DHE (1 μM) fluorescence microscopy 20 min after addition of PDBu (10 μM) or LPS (50 μg/ml) (magnification 200x), (b) DCF-DA (20 μM) fluorescence in a plate reader 20 min after addition of PDBu, or 40 min after addition LPS in the presence or absence of the NADPH oxidase inhibitor VAS2870 (20 μM). (c) Kinetics of ROS formation were measured by L-012 (100 μM) ECL upon addition of PDBu or LPS. Representative images of 2 independent experiments (a) and mean ± SEM of 8 (b) and 8 (c) experiments are shown. versus Ctr (w/o PDBu or LPS) or versus stimulated (with PDBu or LPS) (b) and versus time point 10 min (c).
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