2.6. Cell Culture

RAW264.7 cells were cultured in RPMI-1640 medium (Gibco) supplemented with 10% fetal bovine serum and 1% antibiotics (100 Units/ml penicillin and 100 μg/ml streptomycin) and maintained in a humidified incubator at 37°C under 5% CO₂. When cells were grown to 80% confluence, they were treated with PYR-41 (5 μM, Selleck) for 2 hours, then washed and subjected to ox-LDL (50 μg/ml, Unionbiol) treatment. After incubation with ox-LDL for 12 hours, cells were harvested for RNA extraction or fixed with 4% paraformaldehyde solution for Oil-red O staining.