Cytotoxicity Evaluation of CAME and CAEE

To determine nontoxic concentrations, cells were treated overnight (16–18 h) with 0.1% DMSO or different concentrations of either CAME or CAEE (12.5, 25, 50 and 100 μM). Cell culture media were collected separately and kept on ice. The cells were rinsed with PBS then lysed with 1% Triton X-100 in culture media for 10 min and the lysate were centrifuged at 4 °C for 10 min at 250g. Lactate dehydrogenase (LDH) activity in medium and in lysates was assayed with the LDH-Cytotoxicity Assay Kit II (BioVision, Mountain View, CA) according to the manufacturer's protocol (Smith et al. 2011 (link)). LDH activity was expressed as the ratio of released LDH (medium) to total (medium + lysate) LDH activity.