For cell viability determination Huh-7 and Vero cell monolayers were grown in 96 well plates for 24 h in MEM 5% NBCS. Cell monolayers at 90% confluency were treated with different concentrations of CH223191 (1.25 µM to 80 µM) and kynurenine (5 µM to 320 µM) for 72 h. Mock was treated with the same concentration of DMSO as the highest dose of ligand treatment and was considered as 100% of cell viability. All treatments were performed in quadruplicate. Cell viability was evaluated by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) (Sigma-Aldrich, St. Louis, MO, USA) assay. Absorbance was measured at 570 nm in a FLUOstar OPTIMA microplate reader. Cell viability % was determined by relativizing treatment absorbance results using Mock absorbance values.
Additionally, growth capability and morphology of treated cells were studied under light microscope. Pictures were taken at 72 h with a Nikon Eclipse TS100 microscope; magnification: 100×; NA: 0.25.
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