FIG 5.

The phenotypic effects of pLL on GM-CSF–BMDCs are not due to an impaired switch to glycolytic metabolism. GM-CSF–BMDCs were exposed to pLL, LPS, or both stimuli together for 18 to 22 h; nitrite was measured in supernatants as an indication of ⋅NO output (a) or mitochondrial function (b), and glycolytic fermentative metabolism was assessed with the help of a Seahorse XF-24 extracellular flux analyzer and appropriate inhibitors and/or uncoupling agents (c). Mitochondrial function was assessed by measuring the OCR before and after the sequential addition of the ATP synthase inhibitor oligomycin (oligo), the uncoupler FCCP, and the complex III inhibitor antimycin A (AA). Glycolytic fermentative metabolism was analyzed by measuring the extracellular acidification rate, specifically the portion of this rate that is sensitive to the lactate dehydrogenase inhibitor oxamate and therefore linked to lactate formation and excretion. The graphs show means ± the SD of three to four wells. The results shown are representative of at least two independent experiments.