Proliferation (MTS) assay

Cells were plated in 96-well plates at a concentration of 5×10³ cells/well in complete media. After 24 hours incubation, cells were treated with AB1010, axitinib, indomethacin, or celecoxib (data not shown) in dose-dependent manner alone or in combinations (as specifically shown in the “Results” section) in complete media for an additional 48 hours. Vehicle, dimethyl sulfoxide (DMSO), was used as a control. After treatment, cell viability was measured using MTS CellTiter 96^® Aqueous One Solution Cell Proliferation Assay (Promega Corporation, Fitchburg, WI, USA) according to the manufacturer’s protocol. Briefly, 20 μL of the MTS reagent was added to each well and incubated with cells at 37°C for 1 hour. Absorbance was measured at 490 nM using an FLx800 plate reader (Bio-Tek Instruments, Winooski, VT, USA). The data were shown as mean ± SE of four replicates of three independent experiments and normalized to the DMSO controls.