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. 2020 Sep 7;13:122. doi: 10.1186/s13041-020-00665-7

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© The Author(s) 2020

Open AccessThis 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/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

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Fig. 1 — Comparison of luciferase substrates for in vivo BLI using Bdnf-Luc mice. a. Structure of d-luciferin, seMpai, and TokeOni. b. Schedule of experiments. d-luciferin, seMpai, and TokeOni were administered intraperitoneally to Bdnf-Luc mice, and in vivo BLI was performed 5 min after each administration. Each substrate was injected into mice with at least 2 d intervals. c. Representative images of in vivo BLI using d-luciferin, seMpai, or TokeOni as a luciferase substrate. Bioluminescence; counts indicated by pseudocolored images. Photo; photographs corresponding to bioluminescence images. d. ROI analysis. Data represent the mean ± SEM of four independent experiments using one-way ANOVA with Tukey’s multiple comparisons test (*p < 0.05, **p < 0.01, and ****p < 0.0001). e. Line profiles [counts from (i) to (ii)]. Data represent the mean ± SEM of four independent experiments using two-way ANOVA with Dunnett’s multiple comparisons test [#1; significant difference between d-Luciferin versus TokeOni (p < 0.05), #2; significant difference between d-Luciferin versus seMpai (p < 0.05)]