Pharmacological analysis of AKT activation induced by orbital shaking. (A) MG-63 cells were exposed to orbital shaking for the indicated times in the presence or absence of an inhibitor, and then phosphorylation of AKT was measured by western blotting. (B) Signal intensities of phosphorylated AKT in (A) were normalized to total AKT and plotted as ratios to unshaken (0 min) samples. Open bars, DMSO controls; closed bars, inhibitor-treated samples. (C) MG-63 cells were exposed to orbital shaking or kept static for the indicated times in the presence of the same inhibitor, and then phosphorylation of AKT was measured by western blotting. (D) Signal intensities of phosphorylated AKT in (C) were normalized to total AKT and plotted as shaken/unshaken ratios. Error bars, SD.* p<0.05; **p<0.01. ND, not detected.