Fig. 5.
TGR5 activation promotes beige adipocyte differentiation in vitro. a Heat map showing the expression of Tgr5 and adipocyte and beiging markers from human bone marrow-derived mesenchymal stem cells induced for adipogenic differentiation (GEO Accession number GSE80614)32. Colour key represents row z-score. b mRNA levels of beige remodelling markers Pgc1a, Ucp1, Tbx1, Prdm16, Cidea, Cd137, Pparg2 and Cebpb assessed in the human pre-adipocyte cell line, Simpson Golabi Behmel Syndrome (SGBS). SGBS cells were differentiated in presence or absence of the TGR5 agonist INT-777. n = 6. c mRNA levels of genes described in b in adipocytes differentiated from the stromal vascular fraction (SVF) of TGR5 wild-type (Tgr5+/+) and germline TGR5 knock-out (Tgr5−/−) mice. SVF cells were differentiated for 7 days in presence or absence of the TGR5 agonist INT-777. n = 6. d Representative (n = 6 per group) western blot of PGC-1α, mitochondrial markers (VDAC1 and TOMM40) and beiging markers TBX1 and UCP1 from the cells described in c. PARP1 was used as loading control. e Quantitative densitometry of the western blots showed in d. f Quantification of mitochondrial (16S) vs. nuclear (HK2) DNA ratio from the cells described in c. g Spare respiratory capacity of the cells described in c, calculated as the difference between maximal (FCCP) and basal oxygen consumption rate (OCR). h, i Glycerol (h) and fatty acid (i) release from the cells described in c after 1 h stimulation with the TGR5 agonist INT-777 or vehicle (DMSO). j Basal oxygen consumption rate (OCR) of cells described in c after 3 h pre-incubation with etomoxir and stimulation with the TGR5 agonist INT-777 or vehicle (DMSO). Results represent mean ± SEM. *P ≤ 0.05, **P ≤ 0.01 and ***P ≤ 0.001 vs. Tgr5+/+ cells by one-way ANOVA followed by Bonferroni post hoc test (c, e–j) or Student’s t test (b). Uncropped western blots are provided in Supplementary Fig. 13A–C and Supplementary Fig. 14A