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. 2020 Aug 10;10:13484. doi: 10.1038/s41598-020-70321-6

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© The Author(s) 2020

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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Dosing time-dependent changes in L-OHP pharmacokinetics in mice. (a) Temporal mRNA expression profile of Abcc4 in BMCs of mice. The horizontal bar at the bottom indicates the light and dark cycles. Values are the mean with S.D. (n = 3). There was a significant time-dependent difference in the protein levels of MRP4 (F_5,12 = 4.8254, P = 0.012, ANOVA). (b) Temporal expression profile of MRP4 in BMCs of mice. The left panel shows a representative photograph of Western blotting of MRP4. The horizontal bar at the bottom indicates the light and dark cycles. Values are the mean with S.D. (n = 3). There was a significant time-dependent difference in the protein levels of MRP4 (F_5,12 = 5.626, P = 0.007, ANOVA). Full-size images of western blotting are presented in Supplementary Fig. 5. (c) Time-dependent difference in Pt accumulation in BMCs after treatment with 50 μM L-OHP in the presence or absence of 2 µM ceefourin. Values are the mean with S.D. (n = 4–6). **P < 0.01 significant difference between the two groups (F_3,16 = 12.177, P < 0.001, ANOVA with Tukey–Kramer’s post-hoc test). (d) Time course of serum Pt concentration after a single dose of L-OHP (20 mg/kg, i.v.) at ZT6 or ZT18. Each point represents the mean with S.D. (n = 3). (e) Time course of Pt content in BMCs of mice after a single dose of L-OHP (20 mg/kg, i.v.) at ZT6 or ZT18. Values are the mean with S.D. (n = 3). *P < 0.05, significant difference between the two groups (t₄ = 3.045, P = 0.038, unpaired t-test, two-sided).