miR-206 mediates inhibitory effects of CRTC3 on adaptive thermogenesis. (A) Effect of differentiation mixture (4 d) on RNA amounts for miR-206 in bSVFs prepared from BKO or CRTC3 Tg mice compared with respective control littermates. (B) Occupancy of CRTC3, CREB, and phospho (Ser133)-CREB over a CREB binding site on the miR-206 promoter in WT bSVFs under basal conditions and following exposure to FSK (light blue) for 1 h by manual ChiP assay. (C) Transient assay of HIB1B preadipocytes showing effects of CRTC3 or dominant-negative ACREB expression on WT and cAMP response element (ΔCRE) mutant miR-206 promoter constructs in cells exposed to FSK as indicated. n = 3 per group. (D) Effect of miR-206 on VEGFA secretion from cultured BKO or WT bSVFs infected with adenoassociated virus (AAV)-encoded miR-206 or AAV-GFP control virus. n = 3 per group. (E, Left) Immunoblot showing protein amounts for VEGFA, BDNF, and NGF in BAT from BKO and littermate control mice injected with AAV-miR-206 or AAV-GFP control virus into iBAT. (E, Right) Effect of AAV-miR-206 or control AAV-GFP injection into BAT on core body temperature following 4-h cold exposure in BKO and control mice. n = 4 per each group. Data in A and C represent means ± SD. Data in D and E represent means ± SEM (*P < 0.05; **P < 0.01).