Fig. 3

Increased glycolysis and PPP in 5-FU-R CRC cells. a Major metabolites altered in the glycolytic pathway, the PPP pathway, and the TCA cycle in 5-FU-R CRC cells compared with WT CRC cells. b RT-qPCR analysis of expression of key glycolytic enzyme genes in 5-FU-R cells relative to WT cells. ACTB was used as the internal reference. c RT-qPCR analysis of expression of genes for key enzymes of the PPP pathway in 5-FU-R cells relative to WT cells. ACTB was used as the internal reference. d Western blots of key glycolytic enzymes and G6PD in WT and 5-FU-R CRC cells. β-Actin was used as the internal reference. e 5-FU-R and WT cells were distributed into 96-well Seahorse assay plates and glucose (100 mM), oligomycin (10 μM) and 2-DG (500 mM) were successively added to measure ECAR. f Cell viability was measured by CCK8 assays on treatment with 50 μM 3-BrPA for 48 h. WT or 5-FU-R cells cultured without 3-BrPA were used as a baseline. g Cell viability was measured by CCK8 assays on treatment with 0.5 μM 6-AN for 48 h. WT or 5-FU-R cells cultured without 6-AN were used as a baseline. For all studies n ≥ 3. Data are presented as means ± SEM. Bar chart data were compared by Student’s-t test (* p < 0.05, ** p < 0.01, and *** p < 0.001)