Cytotoxicity (CC50) and inhibition concentration (IC50)

The 50% cytotoxic concentration (CC₅₀) in cells treated with vehicle or with increasing concentrations of MET (0, 1, 2.5, 5, 7.5, 10, 12. 5, 15, 20, and 25 mM) was calculated at 24 h by a colorimetric assay (Roche 11 465 007 001) based on the MTT method ((3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide), according to the manufacturer's instructions using spectrophotometry (BioTek ELx800) measuring absorbance at 540 nm. The Half-maximal (50%) inhibitory concentration (IC₅₀) was calculated using the same concentrations in ZIKV or DENV infected cells. Subsequently, the cells were analyzed by flow cytometry. The CC₅₀ and IC₅₀ of LOV was calculated using the following concentrations: CC₅₀ = 15, 25, 35, 75, 100, 150, 200 and 400 μM; IC₅₀ = 1, 6, 12, 18, 25, 35, 50, 75, 100, 150, 200 μM. The CC₅₀ and IC₅₀ were estimated by linear regression analysis. Three independent experiments in duplicate were performed. The Data were analyzed and plotted with Graph Pad Prism software version 6.0.