Fig. 4: Metabolic tracing confirms potentiation of TH9619-induced thymidylate depletion by hypoxanthine.
From: Formate overflow drives toxic folate trapping in MTHFD1 inhibited cancer cells
a, [U-13C]serine- derived MID of ATP in SW620 WT and MTHFD2−/− cells treated for 24 h with 1 μM TH9619 or TH9975, 50 μM hypoxanthine and 50 μM thymidine (cultured in RPMI-dFBS), means ± s.d. (n = 3); one-way ANOVA with Šídák’s multiple comparisons test for M + 0. P values indicate comparisons to DMSO controls of each cell line. b, ATP levels normalized to PCV in SW620 WT and MTHFD2−/− cells treated for 24 h with 1 μM TH9619 or TH9975, 50 μM hypoxanthine and 50 μM thymidine (cultured in RPMI-dFBS), means ± s.d. (n = 3); one-way ANOVA with Šídák’s multiple comparisons test. c, Relative enrichment of [2,3,3-2H]serine-derived M + 1 to M + 2 isotopologues of dTMP in SW620 WT and MTHFD2−/− cells treated for 24 h with 1 μM TH9619 and TH9975 (cultured in RPMI-dFBS), means ± s.d. (n = 3); one-way ANOVA with Tukey’s multiple comparisons test. d, dTMP levels normalized to PCV in SW620 WT and MTHFD2−/− cells treated for 24 h with 1 μM TH9619 or TH9975, 50 μM hypoxanthine and 50 μM thymidine (cultured in RPMI-dFBS), means ± s.d. (n = 3); one-way ANOVA with Šídák’s multiple comparisons test. e, [U-13C]serine-derived MID of dTMP in SW620 WT and MTHFD2−/− cells treated for 24 h with 1 μM TH9619 or TH9975, 50 μM hypoxanthine and 50 μM thymidine (cultured in RPMI-dFBS), means ± s.d. (n = 3); one-way ANOVA with Šídák’s multiple comparisons test for M + 0. P values indicate comparisons to respective DMSO controls of each cell line. f, dUMP levels normalized to PCV in SW620 WT and MTHFD2−/− cells treated for 24 h with 1 μM TH9619 or TH9975, 50 μM hypoxanthine and 50 μM thymidine (cultured in RPMI-dFBS), means ± s.d. (n = 3); one-way ANOVA with Šídák’s multiple comparisons test. P values indicate comparisons to respective DMSO controls of each cell line.
