Figure 2

Chronic BP3 expression in ob/ob mice by in vivo transfection reduces body weight, WAT, BAT and hepatic steatosis. (A) Schematic diagram showing the protocol for the in vivo transfection with a murine BP3 (mBP3) expression vector in ob/ob mice. Mice received eight treatments and were euthanized 18 days after the first treatment. (B) mBP3 mRNA expression in livers and lungs from transfected ob/ob mice determined by qRT-PCR. Values are shown as fold of control. (C) Western blot analysis for mBP3 protein in lungs. (D) Total body weight and weight gain of control and mBP3 transfected ob/ob mice. Data were analyzed with two-way ANOVA followed by Bonferroni post tests. (E) Representative photographic images of viscera. The white dotted lines indicate visceral WAT. (F) Quantitation of the area occupied by visceral fat. (G) Representative macroscopic photographs of iBAT. (H) iBAT weight. (I) Random blood glucose in fed control and mBP3 transfected ob/ob mice before treatment and at the end of the experiment (day 18). (J) Average Δ glucose in fed control and mBP3 transfected ob/ob mice throughout the course of the experiment. (K) Concentrations of serum metabolites in control and mBP3 transfected ob/ob mice. (L) Representative 4x H&E-stained sections of livers. (M) Semiquantitative scoring of hepatosteatosis from H&E-stained sections of livers. ***P < 0.0001, χ² test. (N) Hepatic lipids and (O) TG levels in control and mBP3 transfected ob/ob mice. (P–R) Liver metabolomics analysis of (P) Glycerol and fatty acids; (Q) Diacylglycerides (DG) and (R) TG isomers in mBP3 transfected ob/ob mice (fold of control). (S) Liver triglyceride levels in control and mBP3 expressing ob/ob and lean C57BL/6 J (WT) mice. All values are expressed as mean ± SEM, n = 3 (control); 6 (mBP3 transfected). ns, non significant; *P < 0.05; **P < 0.01; ***P < 0.0001.