*4.6. Measurement of NO and ROS in RAW 264.7 Cells*

Mouse macrophage RAW 264.7 cells (1 × 106 cells/mL) were cultured in 96 well plates. After overnight culture, cells were pretreated with RI (100, 200. and 400 μg/mL) for 1 h and were then co-treated with LPS (0.5 μg/mL) and RI for 24 h. NO, and ROS activity was measured by Griess reagent and a ROS-Glo H2O2 assay kit according to the manufacturer's recommendation. Intracellular ROS and NO levels were measured at 570 nm by the tunable Versa max microplate reader.
