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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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Analysis of ¹³C-labeled metabolite content derived from [1,2,3-¹³C₃] glucose indicates increased glycolysis and pentose shunt flux in diseased PASMCs, and increased proximate glycolysis metabolites with decreased pentose shunt flux in diseased PAECs. (a) ¹³C₃-Labeled glycolytic intermediates hexose phosphate (HP), fructose bisphosphate (F2P), glyceraldehyde-3-phosphate (G3P), 1,3-bisphosphoglycerate (BPG), phosphoglycerate (PG), phosphoenolpyruvate (PEP), pyruvate (Pyr), and lactate (Lac) in the cell pellet or supernatant (Sup) of control or diseased PASMCs. (b) ¹³C₂-Labeled glycolytic intermediates (derived from pentose shunt metabolism) glyceraldehyde-3-phosphate, 1,3-bisphosphoglycerate, phosphoglycerate, phosphoenolpyruvate, and lactate in in control and diseased PASMCs cell pellet or supernatant; no ¹³C₂-pyruvate was detected. (c) ¹³C-labeled TCA cycle metabolites citrate (Cit), α-ketoglutarate (αKg), succinate (Suc), fumarate (Fum) and malate (Mal); no labeled oxaloacetate was detected. (d) ¹³C₃-Labeled glycolytic intermediates hexose phosphate, fructose bisphosphate, glyceraldehyde-3-phosphate, 1,3-bisphosphoglycerate, phosphoglycerate, phosphoenolpyruvate, pyruvate, and lactate in the cell pellet or supernatant of control or diseased PAECs; no ¹³C₃-glyceraldehyde-3-phosphate or ¹³C₃-phosphoglycerate were detected. (e) ¹³C₂-Labeled 1,3-bisphosphoglycerate, phosphoenolpyruvate, and lactate in in control and diseased PAECs cell pellet or supernatant; no ¹³C₂- glyceraldehyde-3-phosphate, phosphoglycerate, pyruvate or lactate in the pellet was detected. (f) ¹³C-labeled citrate, α-ketoglutarate, succinate, fumarate and malate; no labeled oxaloacetate was detected. (N = 5 samples per group, from 5 different control and 5 different disease subjects; unpaired t-test; *P < 0.05, **P < 0.01; mean ± SEM plotted.) (g) Scatter plot of M + 3 lactate in the supernatant from PAECs and PASMCs obtained from the same individual (N = 3/group).