Fig. 5

PFK15 reduces both ECAR and OCR but does not cause ATP depletion in MIA PaCa-2 cells: a, b: MIA PaCa-2 cells were treated with 10 μM PFK15 or vehicle (DMSO) for 20 min before loading onto a seahorse XFe analyser. Extracellular acidification rate (ECAR, a) and oxygen consumption rate (OCR, b) were recorded after 5 min. MIA PaCa-2 cells were treated for 1 (c) or 5 (d) h with PFKFB3 inhibitor PFK15 (3 or 10 μM), or a cocktail containing 4 μM CCCP, 10 μM oligomycin, 200 μM bromopyruvate and 2 mM iodoacetate as a positive control. ATP was measured using a luciferase-based assay kit and replicates were averaged and normalised to vehicle-treated, time-matched control. Data presented are the mean ± SEM, n = 4 separate experiments, each containing 4–8 replicates. Unpaired t test/ Kruskall-Wallis test; *p < 0.05