Figure 5.

In vitro analysis of the glycolytic state of DU145 and PC3 cell lines. Exponentially growing DU145 and PC3 tumor cells were utilized. A, Cells were exposed to varying concentrations of FX-11, and BrdU uptake was analyzed at 24 hours. B and C, Plated cells were incubated with XF assay medium, and OCR and ECAR were measured. D, Uptake of [2-¹⁴C] pyruvic acid in DU145 and PC3 cells over 20 minutes was determined by assessing cell lysates using a scintillation counter and normalized to protein concentrations. E, Lactate content of exponentially growing tumor cells was measured and normalized to total protein. F, Exponentially growing untreated DU145 and PC3 cells were collected. The expression of the indicated proteins was determined with Western blotting of cell lysates using β-actin as the loading control. G, Cells were exposed to varying concentrations of FX-11, and ECAR was assessed at 140 minutes after FX-11 treatment. H and I, Plated cells were incubated with XF assay medium and treated at the indicated time point (arrow) with pyruvate (8.35 mmol/L) and OCR and ECAR were measured. Bars and points, mean; error bars, SD; ns, not significant; P ≥ 0.05 by paired t test.