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. 2020 Jan 15;10:413. doi: 10.1038/s41598-019-57200-5

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© The Author(s) 2020

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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The TCA cycle of TGF-β-treated PASMCs derives carbons from glucose, glutamine and fatty acids. (a) Effect of the hexokinase inhibitor 2-deoxyglucose (2-DG) on ¹³C₃-labeled glycolysis intermediates hexose phosphate (HP), fructose bisphosphate (F2P), glyceraldehyde-3-phosphate (G3P), phosphoglycerate (PG), phosphoenolpyruvate (PEP), and lactate (Lac) derived from [1,2,3-¹³C₃]glucose in the cell pellet; no labeled ¹³C₃-1,3-bisphosphoglycerate or ¹³C₃-pyruvate were detected. (b) Effect of the transglutaminase inhibitor CB-839 on ¹³C₅,¹⁵N-glutamate (Glm) and ¹³C₅-α-ketoglutarate (αKg) derived from [¹³C₅,¹⁵N₂]glutamine. (c) Effect of the carnitine palmitoyltransferase 1 inhibitor oxfenicine (Oxf) on intracellular fatty acid metabolites derived from ¹³C-labeled fatty acids. (d) Effect of 2-deoxyglucose, CB-839 and oxfenicine on the contribution of ¹³C₂ to citrate deriving from ¹³C-labeled glucose, glutamine and fatty acids, respectively. (e) Effect of 2-deoxyglucose, CB-839 and oxfenicine on the total content of the TCA metabolites citrate (Cit), α-ketoglutarate (αKg), succinate (Suc), fumarate (Fum), malate (Mal) and oxaloacetate (Oxa). (A-D: N = 3 samples per group, E: N = 3 samples per group, with 3 replicates, pooled; unpaired t-test between samples; *P < 0.05, **P < 0.01, ***P < 0.005, ****P < 0.001; mean ± SEM plotted).