Cell viability assay

Cell proliferation was measured by using a cell counting kit-8 (CCK-8) assay (Bestbio, Shanghai, China). Cells were cultured in 96-well plates at a concentration of 5 × 10³–7 × 10³/well. Cells were cultured for 24 h and treated with 0.625, 1.25, 2.5, 5, and 10 μM Amcp. After 24, 48, or 72 h of treatment, the supernatant was completely removed and 100 μL of CCK-8 solution was added to each well, and the cells were cultured for another 2 h at 37 °C. Cell viability was quantified by a SpectraMax M3e instrument (Molecular Devices, San Jose, CA, USA) at 450 nm. Cell viability was calculated for each well using the formula OD₄₅₀ treated cells/OD₄₅₀ control cells × 100%. Assays were performed in three independent experiments.