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. 2012 Dec 13;288(5):3500–3511. doi: 10.1074/jbc.M112.394510

FIGURE 2.

FIGURE 2.

Inhibition of NAMPT by FK866 led to attenuation of glycolysis in cancer cells. A2780 (A and C) and HCT116 (B and D) cells were grown and treated in triplicate with FK866 at various concentrations and ± nicotinic acid (10 μm) as described (“Experimental Procedures”). After the treatment, cells were processed for the analysis of glucose 6-phosphate and fructose 6-phosphate (G6P and F6P), fructose 1,6-bisphosphate (FBP), and glyceraldehyde 3-phosphate and dihydroxyacetone phosphate (G3P and DHAP) (A and B) and 2-phosphoglycerate and 3-phosphoglycerate (2-PG and 3-PG), and phosphoenolpyruvate (PEP) (C and D) levels by LC-MS as described (Experimental Procedures). Because the three pairs of metabolites, glucose 6-phosphate and fructose 6-phosphate, glyceraldehyde 3-phosphate and dihydroxyacetone phosphate, 2-phosphoglycerate and 3-phosphoglycerate, could not be separated chromatographically and they have identical molecular masses, the amount determined by LC-MS represented the total amount of both species in the cell. Error bars represent S.D. *, p ≤ 0.001 versus control.