4.6. LPS Treatment to BV2 Cells for Inflammation Induction

BV2 cells were seeded into a 6-well plate. The number of cells seeded per well of the plate was 2 × 10⁶ cells. Cell starvation was induced by decreasing the concentration of FBS to 0.5%. BV2 cells were then pretreated with different concentrations of the GL extract (0.4, 2, and 10 µg/mL) for 15 min at 37 °C. BV2 cells in each well were cotreated with LPS (1 µg/mL), followed by the incubation at 37 °C for 12 h. One milliliter of supernatant was taken. The cells in each well were washed with cold 1X phosphate buffer solution, followed by the treatment with tissue lysis buffer containing 1% SDS (200 µL). Then, the cell lysate was scraped and collected in an e-tube [65].