Lipid peroxidation assay

To evaluate the malondialdehyde (MDA) content, a Lipid Peroxidation (MDA) Assay kit (cat. no. ab118970; Abcam, Cambridge, UK) was used according to the manufacturer's protocol. A total of 1×10⁶ cells were homogenized on ice in 300 µl MDA Lysis Buffer with 3 µl 100X butylated hydroxytoluene and centrifuged at 13,000 × g and 4°C for 10 min to remove insoluble material. Subsequently, 200 µl supernatant from each homogenized sample was transferred to a microcentrifuge tube and 600 µl TBA solution was added/sample. Following incubation at 95°C for 60 min and cooling to room temperature using an ice bath for 10 min, the 200 µl reaction mixture was transferred into a 96-well microplate for colorimetric analysis. The absorbance was measured at a wavelength of 532 nm and MDA levels were calculated using standard curve analysis according to the manufacturer's protocol.