Additional file 1: Figure S2. of In vitro evaluation of physiologically relevant concentrations of teriflunomide on activation and proliferation of primary rodent microglia

Teriflunomide does not affect cell viability of primary microglia. To investigate possible cytotoxic effects of the drug in the concentrations used in this study, we employed the Alamar blue cell viability assay (Invitrogen, Darmstadt, Germany), which measures the oxidation status of the cells without affecting the function of the electron transport chain. Cells were treated with teriflunomide (stock: 10 mM; dissolved in dimethyl sulfoxide (DMSO, vehicle); 0.25–5 μM) for (A) 24 h, (B) 48 h, or (C) 72 h. The medium was completely changed and replaced with 100 μl cell culture medium supplied with 10 % Alamar blue solution, and cells were further incubated for 4 h. Fluorescence intensity of Alamar blue was measured at 570 nm with a microplate reader (Tecan Sunrise, Crailsheim, Germany). Duplicate measurements were averaged in four independent experiments. Data are normalized to untreated control (1st column), presented as mean ± SD and compared to the untreated control. (TIF 780 kb)