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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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Adequate intracellular nicotinamide adenine dinucleotide (NAD⁺) levels are required to maintain the identity of tumor-initiating cells. (a) U373MG SSEA1⁺ cells were sorted and then incubated with 1 mM of nicotinamide (NAM; NAD⁺ precursor) or 0.1 nM of FK866 (inhibitor of NAD⁺ biosynthesis). Representative cell images at 48 h after treatment are shown. (b) The NAD⁺ levels in each lysate were quantified. (c) Clonogenicity was determined in each condition using limiting dilution analysis. (d) SIRT3 enzyme activity in the mitochondrial fraction and (e) lactate production in each condition were determined. *P<0.05; **P<0.01; ***P<0.001 (Student’s t-test). Scale bar=50 μm.