Figure 3: Increased thermogenic programme in brown adipose tissue by BBR.

(a,b) Representative haematoxylin and eosin staining from interscaptular BAT sections (a) and ratio of BAT to body weight in db/db mice treated with vehicle or BBR (b) (n=5). In a, original magnification, × 200. Scale bar, 100 μm. (c) Mitochondrial density. Representative transmission electronic microscopy images from BAT of db/db mice treated with vehicle or BBR. Scale bar, 10,000 nm in original magnification × 3,400 and 2,000 nm in magnification × 7,400. (d) MtDNA copy number of BAT from BBR-treated and nontreated db/db mice (n=5). (e) Gene expression profiles in BAT. Db/db mice were treated as described in experimental procedure. Real-time PCR analysis of thermogenic and related gene expression is shown (n=5–7). (f) Representative western blots showing the key protein changes in BAT after BBR treatment. (g,h) Fractionated and differentiated primary BAT adipocytes were treated with 2.5 μM BBR for 24 h. Thermogenic gene expression profile after BBR treatment is shown in g, and representative western blot results of the protein expression level is shown in h. Values represent mean±s.e.m. Error bars represent s.e.m., and significant differences compared with vehicle controls are indicated by *P≪0.05, **P≪0.01 and ***P≪0.001 (assessed by Student's t-test).