Prestoblue cell viability reagent 10x solution

Cell viability was estimated using PrestoBlue cell viability reagent 10X solution (Thermo Fisher Scientific, Carlsbad, CA) according to the manufacturer’s instructions. Briefly, TICs in cell culture medium were seeded at 1 × 10⁴ cells/well in a 96-well fibronectin-coated plates treated with vehicle (control) or different doses of LPS and IL-1β for 96 hours and culture medium only for background control. One hour before the end of the culture period, 10X PrestoBlue was added into 96 wells including medium-only wells (blanks) and incubated at 37 °C for 60 minutes. Cell viability was evaluated by fluorescence, which was determined with the use of a microplate reader (FLUOstar Omega, BMG, Durham, NC) with excitation and emission wavelengths of 570 and 610 nm, respectively. Relative cell viability was expressed as a percentage relative to the untreated control cells. This experiment was repeated three times using six to eight replicates for each experiment.