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. 2018 Jan 9;8:153. doi: 10.1038/s41598-017-18530-4

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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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The nuclear-encoded subunit NDUFA9 is imported and assembled efficiently into OXPHOS complex I in 143B MCAD knockout mitochondria. NDUFA9 was radiolabeled by in vitro transcription/translation, followed by incubation for 10 and 60 min with isolated 143B control (CON) or 143B MCAD knockout (KO) mitochondria. (A) SDS-PAGE showing NDUFA9 in its precursor (p) form and as a proteinase K (PK) resistant mature (m) form. NDUFA9 is imported efficiently into both CON and MCAD KO mitochondria in a mitochondrial membrane potential (Δψ_m) dependent manner. (B) BN-PAGE showing the assembly of NDUFA9 into the CI/CIII₂ and CI/CIII₂/CIV supercomplexes (following solubilisation in digitonin, left) or mature complex I (CI, following solubilisation in TX-100, right). (C) Quantitation of NDUFA9 assembly after 60 min of import. There was no difference in the amount of NDUFA9 assembled into the CI/CIII₂/CIV supercomplex (p = 0.07) or complex I (p = 0.14).