Cycloheximide cytotoxicity assay.

The compound 4-(2-hydroxyethyl) piperidine-2,6-dione, also known as cycloheximide, was commercially sourced from Sigma-Aldrich (now Merck), Germany.

Cell culture 96-well vessels were seeded to 4 × 10⁴ cells per well. Triplicate wells of seeded cells were treated with cycloheximide at increasing concentrations. Cells were incubated at 37°C under 5% CO₂ for 24 h and assayed for toxicity using a 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) assay kit (Promega CellTiter 96 Aqueous One Solution Cell Proliferation) according to the manufacturers’ protocol.