Figure 7.

Rapamycin inhibits protein synthesis as well as mTOR and p70S6K in C2C12 cells. The protein synthesis rate was evaluated following treatment by supplementation with puromycin (10 μM) for 30 min in the cell-free supernatant (a, d); the level of total mTOR (b, e) and p70S6K (c, f) following treatment with 1 mM of L-arginine or 1 μM of SNP in the presence of rapamycin (100 nM). When the total protein bands showed significant differences with different treatments, the phosphorylated protein bands were normalized to the total protein bands. In contrast, if the total protein bands were similar across different groups, both the phosphorylated and total protein bands were normalized to β-actin. Data are presented as the means ± SEM (n = 6). ^∗∗P < 0.01 and ^∗P < 0.05 compared with untreated cells. NS, P > 0.05.