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. 2018 Jan 23;8:1395. doi: 10.1038/s41598-018-19836-7

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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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Application of short chain fatty acids (SCFA) failed to change PER2::LUC rhythms of mouse embryonic fibroblasts or cultured PER2::LUC liver slices, with dose and time-of-day dependency. (A) Representative PER2::LUC rhythms of each treatment timing. Circadian time (CT) 0 and CT12 were defined as the bottom and peak of the bioluminescence rhythm, respectively. Vehicle (water) or SCFA + lactate (100 μM) was applied to the media for 30 min (see methods section). Arrows indicate the treatment time and arrow heads indicate the first and second peaks we analysed. (B,C) Dose- or time-dependent phase changes of the first or second peak and amplitude of the first peak after SCFA treatment at CT1, 8, 15, or 22. Value of peak change of the vehicle treatment was set as 0. Value of amplitude of the vehicle treatment was set as 100. All the values are expressed as mean ± SEM (n = 4 in each group, except vehicle at CT22 n = 3). *p < 0.05 vs. vehicle only treatment (Mann Whitney test). (D,E) Representative PER2::LUC rhythms of each treatment timing in cultured liver slices. No difference was seen at the first peak after treatment between the vehicle and SCFA + lactate.