3-(4,5-Dimethylthiazol-2-yl)-5-(3-carboxy-methoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) assay

To investigate the effect of LCH on cell viability, the CellTiter 96™ Assay kit (Promega Corp., Madison, WI, USA) was used, according to the manufacturer's protocol. The HSC2 and HSC3 cells (3×10³ cells/well) were cultured n 96-well plates for 24 h, and treated with LCH for 24 and 48 h at 37°C. Subsequently, 20 µl of MTS solution was added to each well, and the plates were reacted at 37°C for 4 h. The optical density was measured at 490 nm using the Epoch™ microplate spectrophotometer (BioTek Instruments, Inc., Winooski, VT, USA). Cell viability was calculated using the following equation: Optical density ratio of LCH-treated sample/untreated sample ×100 (%).