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. 2018 Jan 16;9:245. doi: 10.1038/s41467-017-02068-0

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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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Fig. 6 — TGR5 activation induces mitochondrial fission in an ERK-DRP1-dependent manner. a, c Representative (n = 6 per group) images of TOMM20 immunofluorescence (in green) on preadipocytes (a) and differentiated adipocytes (c) derived from the stromal vascular fraction (SVF) of TGR5 wild-type (Tgr5^+/+) and germline TGR5 knockout (Tgr5^−/−) mice and stimulated with the TGR5 agonist INT-777 or vehicle (DMSO). Nuclei were stained with DAPI (in blue). Scale bars = 10 μm (a) 25 μm (c). Insets show higher magnification of the reconstructed mitochondrial network (ImageJ program). b, d Quantification of mitochondrial circularity from images as in a and c calculated with ImageJ. n = 6. e, f Representative (n = 6 per group) western blots of TGR5 downstream targets (phospho proteins), their relative controls (CREB, ERK and DRP1) and the mitochondrial protein MFF from the total extract (e) or isolated mitochondria (f) derived from differentiated adipocytes described in c. PARP1 (e) and VDAC1 (f) were used as loading controls. n = 6. Results represent mean ± SEM. **P ≤ 0.01 and ***P ≤ 0.001 vs. Tgr5^+/+ cells by one-way ANOVA followed by Bonferroni post hoc test. Uncropped western blots are provided in Supplementary Fig. 13D and E and Supplementary Fig. 14B–E