Myricitrin Modulates NADPH Oxidase-Dependent ROS Production to Inhibit Endotoxin-Mediated Inflammation by Blocking the JAK/STAT1 and NOX2/p47^phox Pathways

Figure 7

p47^phox and gp91^phox RNAi inhibited the generation of intracellular ROS. (a) RAW264.7 cells were transiently transfected with 1 μg, 2 μg, or 4 μg pFU-GW-p47^phox, pFU-GW-gp91^phox shRNA plasmid, or pFU-GW plasmid vector, respectively. After 48 h transfection, cell lysates were subjected to immunoblotting using antibodies against p47^phox and gp91^phox. GAPDH was used as the loading control. (b) RAW264.7 cells were, respectively, transfected with shRNA-p47^phox and shRNA-gp91^phox or cotransfected with shRNA-p47^phox and shRNA-gp91^phox. In the scrambled RNAi group, cells were transfected with pFU-GW plasmid vector. After 48 h transfection, cells were stimulated with LPS (100 ng/ml) for 5 min. After treatment, cells were washed with PBS and the fluorescence intensity was measured by flow cytometry as described in Materials and Methods. These experiments were independently repeated for three times and the results were expressed as mean ± SD. and versus scrambled RNAi group.

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