Celltiter 96 aqueous one solution cell proliferation assay mts kit

The cytotoxicity of the compounds was also accessed at 10 µM in parallel with viral inhibition screening. Vero cells (1 × 10⁴) were seeded in a 96-well plate and incubated at 37 °C under 5% CO₂ overnight. Compounds were added and incubated for 2 days. DMSO at 1% was used as a mock treatment. Cytotoxicity was measured using the CellTiter 96^® Aqueous One Solution Cell Proliferation Assay (MTS) kit (Promega, Madison, WI, USA) according to the manufacturer’s instructions and analyzed by spectrophotometry at A_{450 nm}.The cytotoxic concentration (CC₅₀) of effective compounds was analyzed by diluting to 8–10 concentrations and adding them to the Vero cells, unless otherwise indicated. Cultures were incubated for 2 days. Cytotoxicity was measured using the CellTiter 96^® Aqueous One Solution Cell Proliferation Assay (MTS) kit (Promega, Madison, WI, USA) according to the manufacturer’s instructions and analyzed by spectrophotometry at A_{450 nm}. The cytotoxicity of each compound was calculated from the nonlinear regression curve, and the concentration required for 50% cell death (CC₅₀) was determined. Results were reported as the mean and standard error of mean (SEM) from three independent experiments.

Cancer cells (1 × 10³/well) were seeded into 96-well plates. After 24 h of growth, cetuximab, AG1478, or Src inhibitor-1 were added at the concentrations indicated, and the cells were allowed to grow for 1 additional day. Cell proliferation was assessed using the CellTiter 96^® Aqueous One Solution Cell Proliferation MTS assay kit (Promega, Tokyo, Japan). Assays were performed in accordance with the manufacturer’s instructions. It has been confirmed that the decrease in MTS values due to cetuximab and AG1478 correlates with the decrease in cell number (Supplementary Figure S1).