![p53 restrains glycolysis and regulates nucleotide metabolism in E1A;HrasG12V MEFs.(A) Schematic representing the flow of glucose-derived carbon through glycolysis and the TCA cycle. Circles indicate carbon molecules present in each metabolite, and filled-in circles (black) represent glucose-derived carbons. (B and C) [U-13C]glucose tracing on sgNTC and sgp53(2) E1A;HrasG12V MEFs in 5% and 21% O2 showing glycolytic (B) and TCA cycle (C) intermediates. Data are mean ± SD of n = 3 cell lines/sgRNA, two independent samples/genotype *, P < 0.05; **, P > 0.005; ****, P < 0.0001; ns, not significant (P > 0.05), one-way ANOVA, Tukey’s multiple comparison test. (D and E) (m+2) citrate:(m+3) lactate ion ratio of sgNTC and sgp53(2) E1A;HrasG12V MEFs cells in 5% (D) or 21% (E) O2. Note different axes. Data represent mean ± SD of n = 3 cell lines/sgRNA, two independent samples/cell line; **, P < 0.005; ns, nonsignificant by Student’s t test. P < 0.05 for sgNTC (5% O2) versus sgNTC (21% O2), one-way ANOVA, Sidak’s multiple comparison test. (F) Schematic of LC-MS/MS analysis of sgNTC and sgp53(2)-targeted E1A;HrasG12V MEFs in 5% O2. Student’s t test (P < 0.05). n = 3 cell lines/sgRNA. (G) Differentially abundant metabolites and the fold change difference of concentration in sgp53(2)-targeted cells relative to sgNTC-targeted cells. Student's t test (P < 0.05). n = 3 cell lines/sgRNA. (H) Pathway enrichment analysis of metabolites exhibiting a more than twofold difference in concentration between sgNTC and sgp53(2)-targeted E1A;HrasG12V MEFs. FDR, false discovery rate.](https://cdn.rupress.org/rup/content_public/journal/jcb/219/11/10.1083_jcb.201908212/4/jcb_201908212_fig5.png?Expires=1771142768&Signature=iHLrA35-lgpJIMFlmGFA8x1DqKSTzE8gSF~r2N4wM68a282bDTcVpQIYcUWl0eFfyLOQnFKy9KtIR53U9zbxkcfME0IBj8FhQkxFbzt5x0k9DvRaIP-Dh8ZMpg7ExucABuGsjRA8xy0qhUQP2am-Xen30xQePQchJCepNKg9NQvjpNB40AQruUE7dRj4wXFR3w4Xn6yrXjiLlqzdxAlsK5C-itgdpl6kh6G-2fOnxoMj2DGV2UU6kelhmx9DU7O7AmwN7ONTA4gkqR3vnjcvW7r9pU-UCi~-NDm~yvtuFktnqbJIsKveLpG2cVz7RAGU0dOzMwcI18LwieoCMCz5PQ__&Key-Pair-Id=APKAIE5G5CRDK6RD3PGA)
p53 restrains glycolysis and regulates nucleotide metabolism in E1A;HrasG12V MEFs. (A) Schematic representing the flow of glucose-derived carbon through glycolysis and the TCA cycle. Circles indicate carbon molecules present in each metabolite, and filled-in circles (black) represent glucose-derived carbons. (B and C) [U-13C]glucose tracing on sgNTC and sgp53(2) E1A;HrasG12V MEFs in 5% and 21% O2 showing glycolytic (B) and TCA cycle (C) intermediates. Data are mean ± SD of n = 3 cell lines/sgRNA, two independent samples/genotype *, P < 0.05; **, P > 0.005; ****, P < 0.0001; ns, not significant (P > 0.05), one-way ANOVA, Tukey’s multiple comparison test. (D and E) (m+2) citrate:(m+3) lactate ion ratio of sgNTC and sgp53(2) E1A;HrasG12V MEFs cells in 5% (D) or 21% (E) O2. Note different axes. Data represent mean ± SD of n = 3 cell lines/sgRNA, two independent samples/cell line; **, P < 0.005; ns, nonsignificant by Student’s t test. P < 0.05 for sgNTC (5% O2) versus sgNTC (21% O2), one-way ANOVA, Sidak’s multiple comparison test. (F) Schematic of LC-MS/MS analysis of sgNTC and sgp53(2)-targeted E1A;HrasG12V MEFs in 5% O2. Student’s t test (P < 0.05). n = 3 cell lines/sgRNA. (G) Differentially abundant metabolites and the fold change difference of concentration in sgp53(2)-targeted cells relative to sgNTC-targeted cells. Student's t test (P < 0.05). n = 3 cell lines/sgRNA. (H) Pathway enrichment analysis of metabolites exhibiting a more than twofold difference in concentration between sgNTC and sgp53(2)-targeted E1A;HrasG12V MEFs. FDR, false discovery rate.
