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. 2016 Mar 25;7(19):27142–27157. doi: 10.18632/oncotarget.8382

Figure 3. Suppression of PFKP inhibits glycolysis, promotes oxygen consumption and increases nucleotide biosynthesis in kidney cancer cells.

Figure 3

A. PFK enzymatic activity of Caki-1 cells transiently transfected with siCtrl, siPFKP1 or siPFKP3. B.-E. Glucose uptake (B), lactate production (C), oxygen consumption rates (D) and ATP levels (E) per cell in Caki-1 cells transiently transfected with siCtrl, siPFKP1 or siPFKP3 at 72 hours after transfection. F. Western blot analysis for Caki-1 cells transiently transfected with siCtrl, siPFKP1 or siPFKP3 using antibodies to p-ACC, ACC, PFKP, p-AMPK, AMPK and tubulin at 72 hours after transfection. G. Real time PCR analysis for Caki-1 cells stably transfected with shRNAs for control (shCtrl) or PFKP (shPFKP1 and shPFKP2). H. ATP levels per cell in Caki-1 cells stably transfected with shCtrl, shPFKP1 or shPFKP2. I. Measurement of glycolytic metabolites in Caki-1 cells stably transfected with shCtrl, shPFKP1 or shPFKP2 using LC-MS. J. Measurement of FBP in Caki-1 cells stably transfected with shCtrl, shPFKP1 or shPFKP2 using an enzymatic method K.-L. Measurement of metabolites in TCA cycle (K) and de novo nucleotide biosynthesis (L) in Caki-1 cells stably transfected with shCtrl, shPFKP1 or shPFKP2 using LC-MS. * indicates p < 0.05 in A-D, G and I-L. The symbol of ns indicates no significant difference in E and H. The following abbreviations are used, fructose 1,6 phosphate (FBP), glyceraldehyde 3-phosphate (G3P), dihydroxy-acetone-phosphate (DHAP), 2-phosphoglyceric acid (2PG), 3-phosphoglyceric acid (3PG), α-ketoglutaric acid (α-KG), ribose 5-phosphate (R5P), 6-phospho-D-gluconate (6PG).