Fig. 4.

Metabolic reprograming in TFAM knockdown colon cancer cells. Control and TFAM knockdown HCT116 cells were labeled with ¹³C₆-glucose (Glc) for 24 h, and levels of metabolites in media and cells were analyzed. a Diagram showing the metabolic network connecting glycolysis with the TCA cycle. The metabolites shown were detected either by NMR in cell media or by IC-MS in cell extracts. b The levels of ¹³C-Glc, ¹³C-Lactate (Lac), and ¹³C-Alanine (Ala) were quantitatively determined by NMR analysis in media. Data represent the mean ± SD (*p < 0.01). c–e The fractional enrichments in the ¹³C isotopologues of metabolites derived from the TCA cycle, including citrate, cis-aconitate, fumarate, malate, aspartate, α-ketoglutarate (α-KG), glutamate and glutathione, as determined by IC-MS analysis. M + 0 (all carbons unlabeled e.g., ¹²C) to M + n isotopologues indicate the number of ¹³C atoms present in each particular metabolite. Data represent the mean ± SD (^¶p < 0.0001, ^§p < 0.001, *p < 0.01, and ^#p < 0.05). All data shown were average measurements from three sets of independently labeled sh-control cells and two sets of sh-TFAM cells. A two-tailed unpaired t-test was used to determine the statistical differences