Figure 5.

Cell non-permeable succinate has no effect on cytokine secretion or cell surface biomarker expression in pro-inflammatory BMDMs. (A–D) BMDMs were pre-treated with disodium succinate (5 mM) 1 h prior to LPS (100 ng/mL) or LPS + IFN-γ (10 ng/mL + 10 U/mL, respectively) stimulation for 24 h. The supernatant was then analyzed for IL-6 (A) and TNF (B) cytokine secretion by ELISA. Additionally, NO secretion (C) was measured by use of Griess Reagent. Cells were subsequently stained with fluorescently labelled antibodies against CD40, iNOS and CD86 (D). Stained cells were then read by a BD LSRFortessa X-20 and cell biomarker surface expression was analyzed in FCSExpress 7. All data are in quadruplicates represented as the mean ± SD and are representative of three independent experiments.