Fig. 6

Effects of adiponectin on AMPK signaling pathway and NF-κB activity in tibialis anterior muscles of mdx mice. The expression of P-AMPK (phosphorylated form) (a) and SIRT1 (b) was analyzed by Western blotting in muscles from the three groups of mice. c Densitometry of immunoprecipitation experiments performed on skeletal muscle lysates, using anti-PGC-1α antibody for precipitation and anti-acetyl-lysine antibody for immunoblotting. d mRNA levels of Myh7, a marker of slow twitch, oxidative myofibers. e mRNA levels of Myh1, a marker of fast twitch, glycolytic myofibers. f mRNA and g protein levels of utrophin A (UTRN) with a representative Western blot and Ponceau S stain. h Immunodetection of NF-κB (p65) in tibialis anterior sections; some positive marked nuclei (brown color) are indicated by arrows. Scale bar = 25 μm. i Quantification of p65 immunolabeling in myofiber nuclei (expressed as percent of total nuclei) in sections (as those shown in g). j Immunoblotting of phosphorylated NF-κB p65 in the same muscles. Levels of P-AMPK, SIRT1, and Acetyl-Lys were normalized to AMPK, actin, and PGC-1α levels, respectively. mRNA levels were normalized to cyclophilin, utrophin A protein levels to Ponceau, and P-p65 to Actin. The subsequent ratios were presented as relative expression compared to WT values. Results are means ± SD; n = 6 mice per group. *p < 0.05; **p < 0.01; ***p < 0.001 vs. WT; ###p < 0.001 vs. mdx mice