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. 2022 Aug 16;82(19):3622–3633. doi: 10.1158/0008-5472.CAN-22-0101

Figure 1.

Figure 1. 2H NMR measurements of labeled fumarate, malate and water in cell culture medium. Representative 2H NMR spectra of medium from untreated S2 cells (A) and from cells 24 hours after the last chemo-radiotherapy treatment (5 Gy per fraction, 15 Gy in total and 50 μmol/L temozolomide), (B) 2 hours after the addition of 5 mmol/L [2,3-2H2]fumarate. Deuterated fumarate, malate and water concentrations in medium from untreated S2 (C), A11 (F), and U87 (I) cells and treated S2 (D), A11 (G), and U87 (J) cells at the indicated times after addition of 5 mmol/L [2,3–2H2]fumarate. Rates of labeled malate production in untreated and treated S2 [***, P = 0.0028; (E)], A11 [*, P = 0.0105; (H)], and U87 [**, P = 0.0075; (K)] cell suspensions (mean ± SD, n = 3 biological replicates). Cell viability was measured in control (L) and chemo-radiotherapy (M) treated S2, A11, and U87 cells at the indicated time points following collection of the media samples.

2H NMR measurements of labeled fumarate, malate, and water in cell culture medium. A and B, Representative 2H NMR spectra of medium from untreated S2 cells (A) and from cells 24 hours after the last chemoradiotherapy treatment (5 Gy per fraction, 15 Gy in total, and 50 μmol/L temozolomide; B), 2 hours after the addition of 5 mmol/L [2,3-2H2]fumarate. Deuterated fumarate, malate, and water concentrations in medium from untreated S2 (C), A11 (F), and U87 (I) cells and treated S2 (D), A11 (G), and U87 (J) cells at the indicated times after addition of 5 mmol/L [2,3–2H2]fumarate. Rates of labeled malate production in untreated and treated S2 (***, P = 0.0028; E), A11 (*, P = 0.0105; H), and U87 (**, P = 0.0075; K) cell suspensions (mean ± SD; n = 3 biological replicates). Cell viability was measured in control (L) and chemoradiotherapy (M)-treated S2, A11, and U87 cells at the indicated time points following collection of the media samples.