Figure 6.

Regulation of the activation of the Erk and Akt signaling pathways by piperlongumine (PL) through reactive oxygen species (ROS) induction. KMH-2 cells were used to evaluate the regulation of signaling pathways under PL treatment. (A) The expression and the activation of Erk, JNK, p38, Akt, and mTOR were examined by western blotting in PL-treated cells. (B) NAC was used to block PL-mediated ROS induction and p38 activation, and the resulting expression of Erk and Akt was examined by western blotting. (C) PD98059 was used to suppress PL-mediated Erk activation, and (D) perifosine was used to promote PL-mediated Akt activation, and the activation of caspase-3 and PARP was examined by western blotting. DMSO was used as a negative control. GAPDH was used as a loading control. Three independent experiments were conducted, and a representative experiment is shown.