2.8. MTT assay for cell viability

Cell viability studies induced by synthesised compounds were evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. RAW264.7 macrophages were seeded in 96-well plates at a density of 5 × 10⁵ cells/mL in complete medium and incubated for 24 h (100 µL/well). Then the cells were treated with different concentrations of synthesised compounds for 24 h. 150 µL MTT (5 g/L in PBS) was added to each well and the cells were further incubated for 4 h. The supernatant was removed and the cells were lysed with 150 µL/well DMSO. The optical density was measured at 570 nm on a microplate reader (Thermo Scientific, MA, USA).