Mts cell proliferation assay kit

In order to obtain half maximal effective concentrations (EC_50s) of cholesterol in MDA-MB-231 and MCF-7 cells, and determine the half maximal inhibitory concentration (IC50) of lovastatin on MDA-MB-231 and MCF-7 cells, proliferation assays with an MTS Cell Proliferation Assay kit (G3582, Promega, Madison, WI, USA) were performed. The MTS assay is based on the reduction of the MTS tetrazolium compound by viable cells to generate a colored formazan dye in metabolically-active cells. According to manufacturer’s instructions, MCF-7 and MDA-MB-231 cells were plated at a density of l0⁴ cells per well in 96 well plates. Cells were treated with varying concentrations of cholesterol, lovastatin, lovastatin (5 μM) + cholesterol, and treatment medium was replaced every 48 h throughout the 5 days’ duration of the experiment. Also, MDA-MB-231 cells were treated with varying concentrations of compound 29, in the absence or presence of cholesterol (5 μM), and with medium replacement every 72 h throughout the 6 day duration of the experiment. MTS reagent (20 μL of per well) was then added to and was incubated for 2 h at 37 °C under standard culture conditions. The optical density (OD) value was measured at 490 nm using a microplate reader (Infinite M200PRO, TECAN).

Cell proliferation assays were performed using a 3-(4,5-diethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-etrazolium, inner salt (MTS) cell proliferation assay kit (Promega) according to the manufacturers’ recommendations. The cells were treated with MTS for 3 h before the termination of HSD17B4 plasmid incubation. The OD readings were performed to determine the number of viable cells at 490 nm. All groups were evaluated in an average of three separate wells per experiment.