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. 2018 Feb 14;8:3029. doi: 10.1038/s41598-018-21050-4

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© The Author(s) 2018

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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Levels of various intracellular metabolites 24 hours after treatment with 0.01% DMSO ± 1 μM GLS1i using the improved culture conditions devised here. A549 and H358 are known as sensitive and resistant cell lines, respectively. Concentration of (a) glutamate and (b) glutamine measured by LC-UV, and relative abundance of the TCA cycle intermediates (measured by LC-MS) (c) alpha-ketoglutarate, (d) citrate and (e) fumarate in cells after treatment with or without 1 μM GLS1i. For each experiment, measurements were performed in triplicate for control and treated conditions. Shown are the mean ± SEM of 3 (A549 cell line; 9 data points per condition) or 2 (H358 cell line; 6 data points per condition) independent experiments. Unadjusted p-values of the differences between control and treated conditions obtained using a two-tailed Student’s t-test, the results of which are denoted with asterisks: **p ≤ 0.01; ***p ≤ 0.001; ****p ≤ 0.0001.