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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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NAMPT inhibition or depletion impairs basal PARP-1 activity. (a) PARP-1, NAMPT and CDA protein levels assessed by immunoblotting in HeLa-Ctrl_(CDA) and HeLa-shCDA cell lines left untreated or treated with 1 μM FK866 for 10 h. Actin was used as protein loading control. (b) Representative immunofluorescence deconvoluted z-projection images of HeLa-Ctrl_(CDA) and HeLa-shCDA cells showing PAR foci in interphase cells. Scale bar: 5 µm. (c) Analysis of PAR foci number in HeLa-Ctrl_(CDA) and HeLa-shCDA cell lines left untreated or treated with 1 μM FK866 for 10 h. The data shown are the means ± SD from four independent experiments (> 350 cells per condition). (d) Representative immunofluorescence deconvoluted z-projection images of PICH-positive UFBs in HeLa-Ctrl_(CDA) and HeLa-shCDA anaphase cells. DNA was visualized by DAPI staining (blue). UFBs were stained with PICH antibody (in green, Alexa Fluor 555). Scale bar: 5 µm. (e) Mean number of UFBs per anaphase cell, for HeLa-Ctrl_(CDA) and HeLa-shCDA cell lines left untreated or treated with 1 μM FK866 for 10 h. The data shown are the means $\pm$ SD from three independent experiments (> 80 anaphase cells per condition). (f) PARP-1, NAMPT and CDA protein levels assessed by immunoblotting in HeLa-Ctrl_(CDA) and HeLa-shCDA cell lines transiently transfected with the indicated siRNAs twice successively for a total of 144 h (96 h + 48 h). HSP90 was used as protein loading control. (g) Analysis of PAR foci number in HeLa-Ctrl_(CDA) and HeLa-shCDA cell lines transiently transfected with the indicated siRNAs. The data shown are the means ± SD from four independent experiments (> 350 cells per condition). (h) Mean number of UFBs per anaphase cell, for HeLa-Ctrl_(CDA) and HeLa-shCDA cell lines transiently transfected with the indicated siRNAs. The data shown are means ± SD from three independent experiments (> 120 anaphase cells per condition). The significance of differences was assessed with Student’s t-test.