Figure 3. Time dependent inhibition of Akt phosphorylation by VCO.

(A, B) Immunoblot analysis of Akt phosphorylation at Thr³⁰⁸ (A) and Ser⁴⁷³ (B) in A549 treated cells with VCO for the indicated time period (upper panel). Fold change represents the protein level of the VCO treated cells relative to the control cells. Bands were quantified by densitometric analysis where pAkt level was then normalized to the total Akt level (lower panel). β-actin served as loading control. (C, D) Immunoblot analysis of pPDK1 Ser ²⁴¹ (C) and mTOR (D) was done at different time hour (0 h, 12 h, 24 h, 36 h) exposure of VCO to A549 cells (upper panel). Bands were quantified by densitometric analysis where pPDK1 or mTOR level was then normalized with β-actin which is represented by folds change (lower panel). Figures are representative of three independent experiments, *p<0.01, **p<0.001 versus control (0 h).