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. 2018 Jun 13;8:9072. doi: 10.1038/s41598-018-27280-w

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© The Author(s) 2018

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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PER2 controls the time-dependent expression of FATP5 in the hepatic membrane fraction. (a,b) Temporal expression profiles of the FATP5 protein in whole-liver lysates (a) and membrane fractions (b) prepared from wild-type mice maintained under a 12-h light/dark cycle condition. The mean peak values of FATP5 protein levels were set to 100. Each value represents the mean with s.e.m. for data from 6–10 mice. There were significant 24-h variations in FATP5 protein levels in the hepatic membrane fraction. (F_5,54 = 28.402; P < 0.001; ANOVA). (c,d) Temporal expression profiles of FATP5 protein in whole-liver lysates (c) and membrane fractions (d) prepared from wild-type and Per2^m/m mice maintained under a 12-h light/dark cycle (left) and constant dark (right) conditions. The mean peak values of FATP5 protein levels from wild-type mice were set to 100. Each value represents the mean with s.e.m. for data from 6 mice. **P < 0.01, *P < 0.05, significant difference between the two groups (F_3,12 = 5.546; P < 0.05; ANOVA with Tukey-Kramer post hoc test, right panel). (e) Co-immunofluorescent staining of FATP5 (green) and NHERF1 (red) in the livers of wild-type and Per2^m/m mice maintained under a 12-h light/dark cycle. Yellow signals indicate the co-localization of FATP5 and NHERF1. Liver sections were prepared at ZT6 and ZT18. Nuclei were also stained with 4′6-diamidino-2-phenylindole (DAPI; blue). Scale bars indicate 10 µm. Data were collected for more than three mice in each group. (f) Temporal profile of the transporting activity of FATP5. Data show time-dependent differences in the area under the curve of the amount of oleic acid uptake into liver slices prepared from wild-type and Per2^m/m mice maintained under a 12-h light/dark cycle. The uptake of oleic acid was assessed for 30 min. Each value represents the mean with s.e.m. for data from 6 mice. **P < 0.01, significant difference between the two groups (F_3,20 = 12.222; P < 0.001; ANOVA with Tukey-Kramer post hoc test). In panels a, b, c, and d, full-size images of immunoblotting are presented in Supplementary Figs 17 and 18.