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. 2020 Aug 17;10:13907. doi: 10.1038/s41598-020-70874-6

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© The Author(s) 2020

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 decrease in basal PARP-1 activity resulting from NAMPT inhibition is independent of NAM accumulation and NAD⁺ levels. (a) and (b) Analysis of intracellular NAM levels in HeLa-Ctrl_(CDA) and HeLa-shCDA cell lines (a) or in BS-BLM and BS-Ctrl_(BLM) cell lines (b) left untreated or treated with 1 μM FK866 for 10 h. The data shown are means ± SD from four (a) or three (b) independent experiments, respectively. (c) and (d) Analysis of intracellular NAD⁺ levels (luciferase assay) in HeLa-Ctrl_(CDA) and HeLa-shCDA cell lines (c) or in BS-BLM and BS-Ctrl_(BLM) (d) left untreated or treated with 1 μM FK866 for 10 h and/or with 500 μM NMN for 24 h. The data shown are means ± SD from five (c) or from three (d) independent experiments, respectively. (e) and (f) Analysis of PAR foci number in HeLa-Ctrl_(CDA) and HeLa-shCDA cell lines (e) or in BS-BLM and BS-Ctrl_(BLM) cell lines (f) left untreated or treated with 1 μM FK866 for 10 h and/or with 500 μM NMN for 24 h. The data shown are means ± SD from four independent experiments (> 400 cells per condition) (e) or from three independent experiments (> 400 cells per condition) (f), respectively. (g) and (h) Mean number of UFBs per anaphase cell, for HeLa-Ctrl_(CDA) and HeLa-shCDA cell lines (g) or for BS-BLM and BS-Ctrl_(BLM) cell lines (h) left untreated or treated with 1 μM FK866 for 10 h and/or with 500 μM NMN for 24 h. The data shown are means ± SD from three independent experiments (> 100 anaphase cells per condition) (g) or from three independent experiments (> 120 anaphase cells per condition) (h). The significance of differences was assessed with Student’s t-test.