Cell Proliferation Assay using MTS

Cell proliferation assays were performed using the CellTiter 96 AQueous One Solution Cell Proliferation Assay (MTS) (Promega, Charbonnières-les-Bains, France), as previously described^{4 (link),7 (link),87 (link)}. Briefly, cells (5 × 10³ cells per well) were plated in triplicate in 96-well plates. After 48 h, cells were incubated with IL-1β and/or TNFα at the indicated concentrations or vehicle only for 48 h with 100 µL culture media (2% charcoal-stripped FBS) (Sigma-Aldrich). Then, 20 µL MTS solution were added to all wells and cells were incubated for 2 h at 37 °C. Absorbance was then read at 490 nm using a Multiskan microplate reader (Thermo Scientific, Illkirch, France). All values were normalized to the values obtained with vehicle-treated cells to control for unwanted sources of variation.