Figure 5

Effect of Lys and/or AICAR treatment on activation of the AMPK, Akt and mTOR pathways, and on the autophagic-lysosomal system in C2C12 myotubes. Cells were treated with DMSO (C), 1 mM AICAR (Ac), 10 mM Leu (L), 10 mM Leu and 1 mM AICAR (AcL), 10 mM Lys (K), or 10 mM Lys and 1 mM AICAR (AcK) for 30 min. The lysates were analyzed for phosphorylation levels of AMPK (a), Akt (b) and 4E-BP1 (c), and the ratio of LC3-II to total LC3 (LC3-I + LC3-II) (d), by Western blotting. Results are expressed as the level relative to the control group. Values are means with SE (n = 3–4). Different letters indicate significant differences among the groups (p < 0.05).