Fig. 4

Lymph node metastases show enhanced PPP and biosynthesis of glutathione and nucleotides relative to the matched primary PDTX in the same mouse. Mice bearing F2 primary PDTX and metastatic lymph node lesions (Met) (from patient UK025) were fed the ¹³C₆-Glc enriched liquid diet for 18 h (cf. Fig. 1) before necropsy. Tissues were processed and analyzed by IC-UHR-FTMS, as described in Fig. 3. Glucose-¹³C is traced through glycolysis, gluconeogenesis (GNG), PPP, the Krebs cycle, ME reactions, and synthetic pathways of GSH, purine nucleotides (PUR), and pyrimidine nucleotides (PYR). Not all possible labeled metabolites were shown. a–l µmole/g protein content of isotopologues of glucose-6-phosphate (G6P), lactate, citrate, αKG, fumarate, malate, Asp, GSH, phosphoribosyl pyrophosphate (PRPP), ATP, UTP, and pyruvate, respectively in primary tumor versus lymph node metastases (cf. Supplementary Fig. 11 for the fractional distribution of these isotopologues). The x-axis represents the number of ¹³C atoms present in each compound; for ATP and UTP, “ring” is the summed value for the ¹³C₆ to ¹³C₉ isotopologues. All other abbreviations and symbols are as in Fig. 3. Values shown are mean ± SEM (n = 5). ^* 0.01 < P < 0.05; ^** 0.001 < P < 0.01; ^*** P < 0.001, two-tailed t-test (see Methods)