Fig 5. Inhibition of AMPK signal reversed the antitumor effects of CAPE in MDA-MB-231 cells.

(A-C) Cells were transfected with siRNA, and the cell viability was determined by CCK8 at 72 h, the colony was investigated by colony formation assay at 5 days and the expression of Cle-caspase3, PARP, and Bcl-xl were detected by western blot at 24 h. (D-G) Cells were treated singly or in combination with Compc (1, 5, 10 μM) and CAPE (50 μM) for 72 h. Cell viability was determined by CCK8. The colony was investigated by colony formation assay at 5 days. The expression of Cle-caspase3, PARP, and Bcl-xl were detected by western blot at 24 h. Apoptosis was determined by Annexin V-FITC/PI dual staining at 48 h. (H-L) Cells were transfected with siRNA or treated in combination with Compc (10 μM) for 24 h, DCFH-DA was used for ROS detection. The lipid peroxidation was determined by C11-BODIPY staining. The expression of GPX4 and Ferritin were detected using western blot. Values represent the mean ± SD from three independent experiments. ***p<0.001: CAPE compared with control, ^#p <0.05, ^##p<0.01: combinatorial Compc+CAPE groups compared with the single CAPE group.