Juglanthraquinone C Induces Intracellular ROS Increase and Apoptosis by Activating the Akt/Foxo Signal Pathway in HCC Cells

Figure 2

JC activates Akt signaling pathway. (a–d) Effects of JC on MAPK and Akt signaling pathways were determined by Western blot analysis. HepG2 cells were treated with either DMSO or 8 μg/mL of JC for different time periods. (e, f) Effects of JC on the Akt signaling pathway were determined by Western blot analysis. HepG2 cells were cultured in a medium without FBS for 12 hours and then treated with either DMSO or 8 μg/mL of JC for the indicated times (e). BEL-7402 cells were treated with either DMSO or 8.7 μg/mL of JC for the indicated times (f). (g) Effects of JC on the NF-κB signaling pathway were determined by Western blot analysis. HepG2 cells were treated with either DMSO or 8 μg/mL of JC for 12 hours and then treated with PMA, which is an activator of NF-κB, for 5, 15, 30, and 60 minutes. Histone H1 was used as a loading control. (h, i) Effects of LY294002 and JC on Akt activation were determined by Western blot analysis. HepG2 cells were pretreated with 50 μM of LY294002 for 1 hour and then treated with 15 nM of rapamycin and 8 μg/mL of JC separately for 3 hours (h). BEL-7402 cells were treated with 30 μM of LY294002 for 1 hour and then treated with 15 nM of rapamycin and 8.7 μg/mL of JC separately for 3 hours (i). GAPDH was used as a loading control.

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