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. 2023 Nov 13;25(12):1774–1786. doi: 10.1038/s41556-023-01280-z

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© The Author(s) 2023

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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Fig. 1 — a, Flow cytometry analysis of EdU incorporation in S-phase cells relative to the non-treated (NT) condition. Cells were treated for 24 h with 10 nM NAMPTi, 2 µM SIRT1i or 10 µM PARPi. The experiment was performed twice with similar results. b, NAD(H) quantification in HeLa and U2OS cells treated as in a. Data are mean + s.d., n = 3 biological replicates, two-tailed Student’s t-test. c, QIBC analysis of γH2AX foci, EdU intensity and DNA content in HeLa and U2OS cells treated as in a. Representative experiment from three independent experiments. d, QIBC analysis of γH2AX foci, EdU intensity and DNA content in HeLa cells treated for 24 h without (NT) or with 10 nM NAMPTi, 10 µM PARPi and/or 2 mM NAD⁺. Representative experiment from three (NT, PARPi and NAD⁺) and two (all other conditions) independent experiments. e, Replication fork speed in HeLa cells treated for 24 h without (NT) or with 10 nM NAMPTi and/or 2 mM NAD⁺. Data are mean ± s.d., means are indicated. Scored forks: NT, 553; NAMPTi, 515; NAD⁺, 552; NAMPTi+NAD⁺, 557. Two-tailed Welch’s t-test. f, Replication fork speed in different cell lines treated for 24 h without or with 2 mM NAD⁺. Data are mean ± s.d., and mean values in kb min⁻¹ of the fork speed are indicated at the bottom of the plot. Scored forks: MRC5 NT, 525; MRC5 NAD⁺, 535; BJ NT, 543; BJ NAD⁺, 523; BJ-T NT, 530; BJ-T NAD⁺, 542; MEF NT, 529; MEF NAD⁺, 612; T98G NT, 519; T98G NAD⁺, 519; U87 NT, 522; U87 NAD⁺, 539; HeLa NT, 543; HeLa NAD⁺, 582. Two-tailed Welch’s t-test. Replication forks were scored for two biological replicates. Numerical data are available in source data files.

Source data