A549 cells increase reliance on the PPP following NAMPT inhibition.

<p>Schematic of key metabolites in glycolysis. B) Cells were treated with 0.4 μM GNE-617 and levels of each metabolite was determined as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0164166#pone.0164166.g002" target="_blank">Fig 2A</a> (Ave ± SD, n = 5). * indicates a p-value of <0.05, and statistical analysis was performed using a Mann Whitney t-test (two-tailed); “ns” indicates not-statistically significant. C) Schematic of key metabolites in the PPP and in nucleotide biosynthesis. D) Cells were treated as in part B and levels of each metabolite was determined as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0164166#pone.0164166.g002" target="_blank">Fig 2A</a> (Ave ± SD, n = 5). * indicates a p-value of <0.05, and statistical analysis was performed using a Mann Whitney t-test (two-tailed); “ns” indicates not-statistically significant. E) Each cell line was exposed to 0.4 μM GNE-617 for 48 hours and labeled with [13C-6]glucose. Levels of newly synthesized pentose-5-phosphate were determined relative to total metabolite levels (Ave ± SD, n = 3). * indicates a p-value of <0.05, and statistical analysis was performed using a Mann Whitney t-test (two-tailed); “ns” indicates not-statistically significant. F) Schematic of the PPP with the step catalyzed by G6PD. Cells were treated with siRNA directed against G6PD and protein levels were examined after 5 days. Full-length western blots are shown in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0164166#pone.0164166.s002" target="_blank">S2C Fig</a>. G) Cells were treated either with siControl or siRNA directed against G6PD for 24 hours, and then treated with different concentrations of GNE-617 for an additional 4 days at which point cell viability was assessed (Ave ± SD, n = 3). * indicates a p-value of <0.05, and statistical analysis was performed using a Mann Whitney t-test (two-tailed); “ns” indicates not-statistically significant.