Fig. 4: Protein and mRNA levels of Lepr, cleaved Caspase-3, Bcl-2 and CCO in the livers of HFD-fed Lepr-WT, Zucker lean, Lepr-KO and ZDF rats, with and without metformin treatment.

A Western blotting for proteins derived from freshly harvested liver tissue. B Quantitative analysis of A. n = 12. Band intensities in A were quantified in Image Lab (Bio-Rad) and the protein expression level was normalized to the β-actin level. Data are presented on a logarithmic scale as the fold change in expression relative to Zucker lean + HFD. C Quantitative real-time RT-PCR analysis for mRNA of the liver tissues (as indicated). n = 12. D Immunohistochemical analysis for Lepr. a Lepr WT + HFD + vehicle; b Lepr WT + HFD + Met; c Zucker lean + HFD + vehicle; d Zucker lean+HFD + Met; e Lepr-KO + HFD + vehicle; f LeprKO+HFD + Met; g ZDF + HFD + vehicle; h ZDF + HFD + Met. Scale bars 30 μm. *p < 0.05 vs. Lepr-KO + HDF + vehicle; #p < 0.05 vs. ZDF + HDF + vehicle.