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. 2023 Oct 2;14:5818. doi: 10.1038/s41467-023-41372-w

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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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 7 — A Spn D39Δcps was treated with NAD⁺ for 9 h or left untreated and bacterial counts were determined. Relative bacterial counts are depicted. B Spn D39 WT, clone 3 and Spn D39Δcps were incubated in a cell culture medium for 6 h with and without NAD⁺ treatment. Afterwards, the intrabacterial ATP was determined by a commercial luminescence assay and normalized against bacterial counts. Measured luminescence is linear to intrabacterial ATP. C, D Intrabacterial ATP after 6 h (C) and bacterial counts after 9 h (D) of combined treatment of Spn D39 WT with 500 µM NAD⁺ and 5 mM pyruvate. Statistics: two-tailed t-test (A, B); Two-way-ANOVA with Fisher’s LSD (C, D); significance was determined against uninfected/untreated controls if not indicated otherwise; N = 4 biologically distinct samples; box plots: line at mean; box ranges from min to max; results are normalized against untreated controls. WT wild type.