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. 2019 Oct 1;9:14132. doi: 10.1038/s41598-019-50661-8

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© The Author(s) 2019

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Decreased glutamine levels contribute to reduced GSH in TRPM2 depleted cells. (a) Schema of glutamine metabolism and GSH production. (b–d) Metabolomic quantitation of TRPM2 depleted and scrambled control cells treated with or without doxorubicin (n = 6 replicates/group) showed decreased (b) ATP and GTP, (c) glutamine and GSH, and (d) NAD⁺ and NADP⁺ in KO cells compared to scrambled control. *p ≤ 0.003, group effect; **p < 0.005, group x doxorubicin exposure time interaction effect, two way ANOVA. (e) Western blotting of proteins involved in GSH and α-ketoglutarate synthesis including GLS, GCLC, GCLM, GSS, GLUD, c-Myc, xCT, and β-actin. Representative blots from three experiments are shown. Densitometry measurements were normalized to the average of each blots’ untreated scrambled controls and mean densitometry measurements from the three experiments are shown in (f). p values analyzed with two-way ANOVA: GLS and GSS (*p < 0.0001), GCLC (*p < 0.007), GCLM (*p < 0.003), GLUD (*p < 0.04), xCT (*p < 0.05), group effect; GCLC (**p = 0.006), c-Myc (**p = 0.002), group x doxorubicin exposure time interaction effect.