Phosphorylation of Akt induced by Angelica Sinensis (AS). (A) Upper panel showed a representative result of western blot analysis of total- Akt (t-Akt) and phosphor-Akt (p-Akt) 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) Akt phosphorylation level at 15 min in wortmannin-treated myotubes. (C) Akt phosphorylation level at 45 min in wortmannin-treated myotubes. p-Akt and t-Akt were normalized by individual β-actin. The results of the densitometric analysis of the western blot membranes [upper panels in (A), (B) and (C)] were depicted in the lower panels as the ratio of p-Akt against the t-Akt signal (mean ± SD, n = 3), respectively. Vertical axis represented relative p-Akt level compared with pre-treated myotubes (A), or non-treated myotubes (B) and (C). 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) and (C). *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) and (C) (Scheffe’s post hoc analysis, P < 0.05). #Significant inhibitor effect in the same group (Scheffe’s post hoc analysis, P < 0.05).