Fig. 3

Effects of SH003 on the activation of intracellular signaling pathways and induction of apoptosis by JNK phosphorylation in DU145 cells. DU145 cells were exposed to 50, 250, or 500 μg/mL SH003 for 15 min and western blotting was used to determine expression levels of a ERK, JNK, and p38 related MAPK components, b SRC-STAT3 signaling pathway components, or c PI3K-AKT pathway proteins. β-actin served as the internal control. d Cells were pretreated with 10 μM SP600125 for 30 min and then treated with 500 μg/mL SH003 for 24 h. Levels of apoptosis-related proteins and p-JNK were measured by western blotting. e Cells were pretreated with 10 μM of SP600125 for 30 min and then treated with 500 μg/mL SH003 for 48 h before staining with Annexin V and 7-AAD at room temperature in the dark. Levels of apoptosis were analyzed by flow cytometry. *P < 0.05