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Results: 4

1.
Figure 2

Figure 2. 13C labeling of selected metabolites in myc−/− and myc+/+ cells. From: c-Myc activates multiple metabolic networks to generate substrates for cell cycle entry.

13C enrichment at individual carbons (left). Data is calculated from integrated peak areas relative to internal standard. Mean of 3 experiments. Significant differences (*=p<0.05, **=p<0.001) assessed by Student’s t test. Corresponding NMR spectra (right) for (A) lactate, (B) alanine, (C) glycine, (D) NXP ribose C4’ and adenine C2.

Fionnuala Morrish, et al. Oncogene. ;28(27):2485-2491.
2.
Figure 4

Figure 4. From: c-Myc activates multiple metabolic networks to generate substrates for cell cycle entry.

Myc regulation of glucose metabolism may provide key intermediates, energy and reducing power for cell proliferation. 13C-labeled metabolites from this study are boxed. Dashed arrows link mitochondrial metabolites with reversible (double-ended arrows) or irreversible (single arrows) reactions. Both glycolysis and the mitochondrial TCA cycle generate metabolic intermediates, in addition to ATP, providing building blocks for protein, lipid and nucleic acid synthesis. Post-translational protein modification may be subject to substrate-level control, including extra-mitochondrial acetyl-CoA, derived from intra-mitochondrial pyruvate and fatty acid metabolism, and glucosamine-6-phosphate, derived from fructose-6-phosphate and glutamine.

Fionnuala Morrish, et al. Oncogene. ;28(27):2485-2491.
3.
Figure 3

Figure 3. Metabolic pathways required for cell cycle progression in myc+/+ cells. From: c-Myc activates multiple metabolic networks to generate substrates for cell cycle entry.

(A) Proton spectra for phosphocholine (3.2 ppm). (B) Immunoblots showing O-GlcNAcylated proteins and OGT expression with serum stimulation. Cyclin E and tubulin are positive and loading controls. Densitometry values indicated below each lane (ImageQuant). (C) Dose-response assay of cell growth by HO33342 staining for methotrexate and DON. TGR represents myc+/+ cells. Relative growth rates of untreated cell lines: myc−/− 0.69, mycER 0.83. Representative of 3 experiments in triplicate.

Fionnuala Morrish, et al. Oncogene. ;28(27):2485-2491.
4.
Figure 1

Figure 1. 13C NMR analysis of 13C glucose metabolism in myc−/− and myc+/+ cells during cell cycle entry. From: c-Myc activates multiple metabolic networks to generate substrates for cell cycle entry.

(A) 1H decoupled 13C spectra. Key. (1) C3 alanine, (2) C3 lactate, (3) C3 glutamate, (4) C4 glutamate, (5) C2 glycine, (6) C5 proline, (7) C2 glutamate, (8) C2 lactate, (9) ribose and sugar moieties, (10) C4‘ NXP, (11) C2 Adenine, (12) C1 glutamate, (13) C5 glutamate, (14) C1 lactate. TMPSA is internal standard. NXP refers to the mono- di- or triphosphate form of any nucleotide. (B) tcaCALC-derived relative fluxes. (C) Percentage contribution of 13C labeled glutamate to total glutamate by LC/MS/MS mass isotopomer analysis.

Fionnuala Morrish, et al. Oncogene. ;28(27):2485-2491.

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