2.5 RAW 264.7 cells culture and treatment
RAW 264.7 cells were purchased from Cell Bank, and cultured in HG-DMEM medium containing 10% FBS. All cells were seeded into 24-well cell culture plates at a concentration of approximately 3x105 cells per well, maintained in a humidified atmosphere of 5% CO2 at 37 ˚C, and allowed to grow for 24 h. LPS was used to trigger inflammatory responses. RAW 264.7 macrophages were treated with LPS alone (100 ng/mL) or LPS with 270 for 24 h, harvested cells were used for RNA extraction. RAW 264.7 cells were pretreated with 270 for 4 h before LPS (100 ng/mL) treatment for 30 min, and cells lysates were obtained for western blot.