Fig. 3.

The involvement of oxidative stress and nuclear transcription factor NF-κB activation in the induction of PAI-1 in VSMCs stimulated with 10 µM lysoPC. (A) Effects of vitamin E (50 µM), a potent antioxidant, or CAPE (20 nM), an inhibitor of NF-κB, on PAI-1 expression induced by lysoPC for 6 hr as assessed by Western blot analysis. Representative blots are shown in the upper section of each panel. The intensity of the bands was densitometrically determined and normalized to that of corresponding β-actin bands. Graphic data are depicted in the lower part of the panel. (B) Intracellular ROS production detected by DCF fluorescence under confocal microscopy (× 100). (C) Kinetics of intracellular ROS production as assessed by flow cytometry analysis using DCF-DA. (D) Effects of lysoPC for 1 hr on NF-κB-mediated transcriptional activity as measured by luciferase reporter assay. ^*P < 0.05 vs baseline; ^†P < 0.05 vs DSF medium; ^‡P < 0.05 vs vehicle-treated control.