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. 2023 Jun 8;19(9):1147–1157. doi: 10.1038/s41589-023-01350-1

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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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Extended Data Fig. 10 — a, b. BRET-FRET/eGFP ratios of ChemoL-NAD_CPY expressed in the nucleus (a) or the mitochondria (b) of U-2 OS cells upon treatment for 24 h with DMSO (Ctrl), 1 mM NR, 100 nM FK866 or 100 nM FK866 plus 1 mM NR. Represented are the BRET-FRET/eGFP ratios of single wells (circle) and means ±s.d. (black line) (n = 6 wells from a single experiment). p-values are given based on unpaired two-tailed t-test with Welch’s correction (**** p < 0.0001, ** p = 0.0041, ns p = 0.5718). c, e. Luminescent intensity (LI) spectra of ChemoL-CaM (c) or ChemoL-ATP (e) at different concentrations of calcium and ATP, respectively. The spectra were normalized to the isosbestic point at 593 nm. Constructs were labeled with CPY. Shown are the means of 3 technical replicates. d, f. Analyte titrations of ChemoL-CaM (d) and ChemoL-ATP (f) labelled with CPY. Shown are the mean ±s.d. of BRET-FRET/eGFP ratios (n = 3 technical replicates). Indicated are also the maximum BRET-FRET/eGFP ratio changes (^MaxΔR/R₀).

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