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. 2019 Oct 21;9:15047. doi: 10.1038/s41598-019-51517-x

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© The Author(s) 2019

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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Mitochondrial (mt)DNA depletion and mitochondrial dysfunction in A549 Rho-0 cells. A549 wild-type (wt) and Rho-0 cells were grown to confluence and serum deprived for 24 hrs. (A) Cells were stained with Picogreen for detection of the DNA and Mitotracker DeepRed. Co-localization of DNA and mitochondrial bodies indicates mtDNA. Representatives of 3 independent experiments are shown. (B) Cells were stained with JC1 staining mitochondrial membrane potential is lowered by depletion of mtDNA in the A549 Rho-0 cells when compared to control cells. (C) ATP (mean ± SEM, n = 6–7) and (D) lactate (mean ± SEM, n = 3) were measured in cell-free culture supernatants. (E) RNA was isolated, cDNA synthesized, mRNA expression of NADH dehydrogenase (complex I), cytochrome c oxidase (complex IV) subunit III and ATPase subunit F1α (complex V) analyzed by qPCR and related to the expression of housekeeping genes β2µG and PPIA. (F) Cell lysates were prepared and complex III, Complex IV and the ATPase subunit components were detected by western blotting, analysed by densitometry and related to GAPDH (mean ± SEM, n = 3). *p < 0.05 and **p < 0.01 between the indicated values as measured by the one-tailed Mann-Whitney test.