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. 2017 Jun 9;49(6):e344. doi: 10.1038/emm.2017.74

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Copyright © 2017 The Author(s)

This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivs 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by-nc-nd/4.0/

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The effect of nicotinamide nucleotide transhydrogenase (NNT) overexpression or knockdown in SSEA1⁺ tumor-initiating cells is shown. Mitochondria were fractionated from retroviral NNT-overexpressed or siNNT-transfected U373MG SSEA1⁺ cells, and (a) nicotinamide adenine dinucleotide (NAD⁺) levels and (b) SIRT3 enzyme activity were determined at 3 days after treatment. (c) Lactate production was examined in the total lysate in each condition. (d) Each condition at 3 days after culture, the oxygen consumption was measured in over 80 min using an oxygen probe and (e) the ATP concentration in cells of each group was determined. (f) Immunocytochemistry of each condition was fixed at 7 days after treatment. Antibodies used in each condition are shown on the left. 4,6-Diamidino-2-phenylindole (DAPI) staining (blue) was used to identify nuclei.