Fig. 4.
Effects of dietary leucine on SIRT1 signaling and mitochondrial biogenesis in brown adipose tissue (BAT). A: BAT homogenates were subjected to WB analysis using an anti-SIRT1 antibody, total RNA was extracted from BAT, and mRNA levels of SIRT1 were analyzed by quantitative real-time PCR (n = 5). *P < 0.05 vs. ND or HFD. B: BAT homogenates were subjected to WB analysis using an anti-NAMPT antibody (n = 5). *P < 0.05 vs. ND; †P < 0.05 vs. HFD. C and D: PGC1α or FoxO1 was immunoprecipitated (IP), and WB was performed using indicated antibodies. E: expression of genes related to mitochondrial biogenesis in BAT. F: mtDNA/nuclear DNA ratio in BAT was determined by quantitative real-time PCR. G: BAT CS activity. H: ATP levels in BAT. I: expression of mRNA controlling FAO (n = 5). *P < 0.05 vs. HFD. J: BAT was formalin fixed and embedded in paraffin; sections were stained with hematoxylin and eosin. Three samples in each group were analyzed and showed similar results.