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. 2018 Mar 23;8:5118. doi: 10.1038/s41598-018-23175-y

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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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Downregulation of the components of Fe-S cluster biogenesis machinery in Irp1 and 2 deficient fibroblasts is specifically associated with impaired electron transport chain (ETC). (a) A representative graph of in-gel assays of mitochondrial (m-Aco, encoded by Aco2) and cytosolic (c-Aco, encoded by Irp1) aconitases in Irp deficient MEF cells (upper panel). The protein levels of Aco2, Irp1, and cytosolic Fe-S containing enzyme xanthine oxidative (Xod) were detected with western blotting. (b) The activities of Xod, m-Aco, and c-Aco were quantified. (c) Activities of ETC complexes were determined in Irp deficient MEFs. (d) Western blot analysis of mitochondrial proteins including Ndufs3 (a subunit of CI), SdhA and SdhB (subunits of CII), Uqcrc1 and Uqcrfs1 (subunits of CIII), Fech (matrix protein), CytC (intermembrane space protein), Vdac (outer membrane protein), and citrate synthase (Cs, matrix non-Fe-S protein). A representative image set is presented. (e) Activities of citrate synthase, which is a mitochondrial non-Fe-S enzyme, were determined. CI/CII/CIII/CIV: Complex I/II/III/IV. Values represent mean ± SEM (n = 10 for (e), n = 3, each duplicates for (b) and (c)). A one-way ANOVA was performed. *p < 0.05, **p < 0.01, ***p < 0.001 compared with WT.