4.2. BV-2 Microglial-Cell Culture

The BV2 microglia cells were acquired as described previously [35,36]. The BV-2 microglial cells were cultured in DMEM supplemented with 5% FBS and 1% of 100 units/mL of penicillin/streptomycin at 37 °C in a humidified 5%-CO₂ incubator. In all of the experiments, the cells were seeded at a density of 2.5 × 10⁵ cells/mL, and they were 6 h after then pretreated for 1 h with different concentrations of scoparone (100 μM, 250 μM, and 500 μM), followed by an incubation with LPS (200 ng/mL) for the indicated times (1 h, 6 h, and 24 h). In the cell morphology experiment, the BV2 microglial cells were pretreated with 500 μM of scoparone followed by LPS (200 ng/mL) induction for 24 h, and the cellular morphology images were observed by using phase contrast microscopy (Axio; Carl Zeiss, Jena, Germany).