Figure 4

Phosphorylation of mTOR induced by Angelica Sinensis (AS). (A) Upper panel showed a representative result of western blot analysis of total (t-mTOR) and phosphor-mTOR (p-mTOR) levels in the myotubes treated with 10 ng/mL of IGF-1 in 2% HS/DMEM for 45 min, or AS (10 ng/mL of AS in 2% HS/DMEM) for 5 to 60 min. (B) mTOR phosphorylation level at 30 min in wortmannin-treated myotubes. p-mTOR and t-mTOR were normalized by individual β-actin. The results of the densitometric analysis of the western blot membranes [upper panels in (A) and (B)] were depicted in the lower panels as the ratio of p-mTOR against the t-mTOR signal (mean ± SD, n = 3), respectively. Vertical axis represented relative p-mTOR level compared with pre-treated myotubes (A), or non-treated myotubes (B). Data were analyzed with one-way ANOVA with time factors in (A). Data were analyzed with two-way ANOVA with group and inhibitor treat as factors in (B). *Significant time effect compared with pre-treat in (A) (Scheffe’s post hoc analysis, P < 0.05). *Significantly different compared with the NON without inhibitor wortmannin in (B) (Scheffe’s post hoc analysis, P < 0.05). #Significant inhibitor effect in the same group (Scheffe’s post hoc analysis, P < 0.05).