The optimized assay of maximum glycolytic capacity.

<p>Raw traces of (<i>a</i>) oxygen consumption rate (OCR) and (<i>b</i>) extracellular acidification rate (ECAR) in HEK293 fibroblasts after sequential additions of 10 mM glucose, 1 μM rotenone plus 1 μM myxothiazol, and 200 μM myxothiazol plus 1μM FCCP. <i>c</i>: Respiratory (open column sections) and glycolytic (blue column sections) proton production rates (PPR) of the experiment exemplified in A and B calculated using <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0152016#pone.0152016.e001" target="_blank">Eq 1</a>. Shaded wedges indicate glycolysis under basal conditions (lightest), ATP demand-limited glycolytic rate (medium), and maximum glycolytic capacity (darkest), with the difference between the basal rate and the maximum glycolytic capacity defined as the glycolytic reserve; Data are means ± SEM of n = 4 independent biological replicates.