Cell Culture and Viability Assays

RAW 264.7 and BMM cells (ATCC, Manassas, VA, USA) were cultured in alpha-MEM supplemented with 10% heat-inactivated FBS, 2 mM L-glutamine, and 100 U/ml penicillin/streptomycin. Incubations were carried out at 37°C in 5% CO₂. We performed an MTT assay following the manufacturer’s instructions to detect CTU cytotoxicity. RAW264.7 and BMM cells were seeded on a 96-well plate at a concentration of 5 × 10³ cells/ml, followed by treatment with different concentrations and incubation for 120 h. The concentration of MTT solution was 50 mg/ml, and the absorbance at 490 nm was measured by an ELISA plate reader (Männedorf, Swiss).