Fig. 3

Limitation of using TokeOni for repeated in vivo BLI. a. Schedule of experiments. In vivo BLI was performed before (i) and after (ii) the intraperitoneal administration of each substrate to Bdnf-Luc mice (First injection). Six hours after the first injection, in vivo BLI was performed before (iii) and after (iv) substrate administration again (Second injection). b. ROI analysis to measure counts at each step. d-luciferin was administered according to the schedule shown in Fig. 3a. Data represent the mean ± SEM of three independent experiments using one-way ANOVA with Tukey’s multiple comparisons test (***p < 0.001, NS; not significant). c. ROI analysis. TokeOni administration and in vivo BLI were performed according to the schedule shown in Fig. 3a. Data represent the mean ± SEM of four independent experiments using one-way ANOVA with Tukey’s multiple comparisons test (**p < 0.01, ***p < 0.001, NS; not significant). d. Schedule of experiments. In vivo BLI was performed 5 min after the intraperitoneal administration of TokeOni to Bdnf-Luc mice (0 h). Then, in vivo BLI was repeatedly performed at 3, 6, 9, 12, and 24 h without further TokeOni administration. Twenty-four hours after in vivo BLI, TokeOni was re-injected into the mice, and in vivo BLI was performed again (v). e. Representative images of in vivo BLI. The pseudocolored range shown to the left (from 0.5 × 10³ to 3.0 × 10³) corresponds to the images at (0 h) and (v), and the range shown to the right (from 0.2 × 10³ to 1.0 × 10³) corresponds to the other images. f. ROI analysis. TokeOni administration and in vivo BLI were performed according to the schedule shown in Fig. 3d. Data represent the mean ± SEM of four independent experiments using one-way ANOVA with Tukey’s multiple comparisons test (****p < 0.0001, NS; not significant)