Tivantinib

The cell proliferation assay was performed in triplicate with a cell density of 4 × 10³ cells/well in 96 well plates and eight 3X serial dilutions of the drug, ranging from 100 to 0.046 μM. Two MET tyrosine kinase inhibitors, crizotinib and tivantinib, were purchased from MedChemExpress (NJ, USA). Serial dilutions of Curcumin (Sigma-Aldrich, US) ranging from 50 to 0.781 μg/ml were used as standards to ensure the consistency of the experiments across replicates. Cells were incubated at 37°C for 72 hours in complete media and cell viability was assessed using the CellTiter Aqueous Non-Radioactive Cell Proliferation Assay, (Promega, Madison, WI, USA), according to the manufacturer’s protocol. The absorbance at a wavelength of 490 nm was measured using the Epoch Microplate Spectrophotometer (BioTek Instruments, Inc., US). The half-maximum inhibitory concentration (IC50) was assessed using Gen5 software, version 1.11.5. This program calculates the IC50 using the dose-response equation [Y = {(A – D)/[l + (X/C)B ]} + D], where X is the drug concentration, Y is the absorbance at 490 nm, A is the upper asymptote, B is the slope, C is the IC50, and D is the lower asymptote.