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. 2017 Oct 20;7:13663. doi: 10.1038/s41598-017-13961-5

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© The Author(s) 2017

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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ATP production in NHM and VHM in basal and forced mitochondrial activity. (A) Quantification has been performed by fluorimetric kit. It was carried out in basal condition (standard culture medium) and after 12 days treatment with 20 mM L-glutamine. Data are mean ± SD of all the performed experiments. *p = 0.008; **p = 0.009. (B) Flow cytometric analysis of MFI for HKII, PKM2, and PDHK1. (C) The percentage of positive cells for HKII and PKM2 was also evaluated and an increased expression was observed in VHM in basal condition; NHM down-regulate its expression after L-glutamine supplementation whereas VHM aren’t affected by the presence of the alternative substrate, underlying the difference between the two populations. (D) The activity of HKII was also forced in a compensatory strategy. Data are mean ± SD of all the performed experiments. *p = 0.05; **p = 0.01. (E) A relevant proton leak was detected in VHM, underlying the inability of ETC to correctly finalize the specific activity. Consequently, the mitochondrial performance, represented by BHI, was negatively affected. BHI is calculated as (ATP × Reserve Capacity)/(proton leak × non mitochondrial respiration). Data are mean ± SD of all the performed experiments. *p = 0.01.