FIGURE 3.

Ffar4 agonist recapitulates EPA-mediated brown thermogenesis. Brown progenitor cells were isolated from iBAT and induced differentiation for 7 days with an increasing dose (0, 1, 10, and 20 μm) of GW9508, a FFAR4 agonist. A, brown signature gene expression profile of Ucp1, Prdm16, Cidea, Elovl3, Glut4, and Rip140 by qPCR. B, changes of miRNA expression of miR-30b, miR-193b, miR-365, miR-378, miR-196a, miR-27a, miR-93, miR-106b, miR-133, and miR-155. C, representative Western blotting analysis of Ucp1, Pparγ, aP2, and β-actin. D, OCR in brown adipocytes grown with vehicle (Veh) (green) or 20 μm GW9508 (blue) determined by Seahorse extracellular analyzer. Arrow indicates the addition of respiratory inhibitors of oligomycin (Oligo), FCCP, and antimycin A plus rotenone (A + R). All data are presented as mean ± S.E. A, values not sharing a common letter differ significantly (p < 0.05) by one-way ANOVA (n = 5 per group). B (n = 5); D (n = 6), *, p < 0.05; **, p < 0.01; ****, p < 0.0001 by Student's t test.