Fig. 6
β-LAP inhibited ROS production via suppression of NADPH oxidase subunits. a BV2 cells or b primary microglia were pretreated with β-LAP (0.5, 1, and 2 μM, for 1 h), followed by LPS (100 or 10 ng/ml, for 16 h), and stained with 50 μM H2DCF-DA. DCF fluorescence intensities were measured using a microplate fluorometer. The data are expressed as the means ± SEM of three independent experiments. *P < 0.05, significantly different from the LPS-treated group. c A representative confocal image of DCF-derived fluorescence (green) in BV2 cells (n = 3), with an original magnification of ×200. d RT-PCR analysis for NADPH oxidase subunits (p47phox, p67phox, gp91phox, p22phox). BV2 cells were pretreated with β-LAP (0.5, 1, and 2 μM, for 1 h) followed by LPS (100 ng/ml), and total RNA was isolated at 2 h after LPS treatment. Representative gels are shown in the left panel, and quantification data are shown in the right panel (n = 3). e Western blot analysis for phosphorylation of the p47phox subunit (n = 3). BV2 cells were pretreated with β-LAP (0.5, 1, and 2 μM, for 1 h), followed by LPS (100 ng/ml, for 30 min), and then subjected to immunoblot analysis using antibodies against phospho-p47phox. Quantification data are shown in the graph. *P < 0.05, significantly different from the LPS-treated group