Figure 6.

SMEF inhibits RANKL-induced reactive oxygen species (ROS) production using cellular dichlorodihydrofluorescein diacetate (DCF-DA) assay. RAW 264.7 cells were seeded in a 96-well plate, pretreated with the SMEF (50 µg/mL) for 24 h, and stimulated by RANKL (100 ng/mL) for 1 h. Then, 40 µM of DCFH-DA solution was added for 30 min. Treated cells were measured with a fluorescent microplate reader. Then, 50 µM N-acetylcysteine (NAC) and 250 µM ascorbic acid (Vit C) were used as a positive control. All data are presented as the mean ± SD of three independent experiments. ^## p < 0.001 compared with the untreated control (CON); ** p < 0.001 compared with RANKL treatment (RANKL).