Cell Viability Assay with CCK-8

The CCK-8 detection kit (Beyotime, China) was used to investigate the effects of different compounds on cell viability, as described previously (7 (link)). Briefly, approximately 5 × 10³ GCO cells were seeded into 96-well plates. Cells were treated with compounds at different concentrations for 24 h. Then, 10 μL of CCK-8 solution was added to each well and incubated at 28°C for 4 h, and the absorbance at 450 nm was measured using a microplate reader (Bio-Rad, USA). The untreated cells were considered the positive control, while the wells containing no cells and only culture medium were used as blank controls. Data are presented as the means ± SD of three replicates.