Fig. 6

Cirsiliol exerts its action through the AKT/FOXO1 axis. A Molecular docking model of cirsiliol versus AKT. B Western blot assays were performed on lysates from U2OS and U2OS/MTX cells treated with cirsiliol. The antibodies used included AKT, p-AKT, FOXO1, p-FOXO1, and ACTB. C SC79 and siFOXO1 were added to detect the expression levels of AKT, p-AKT, FOXO1, p-FOXO1, and ACTB. D Incidence of cell apoptosis in groups DMSO, cirsiliol (20 µM), cirsiliol + SC79, and cirsiliol + siFOXO1. E Statistical graphs done for D. Three replicate experiments were performed. F, G Immunofluorescence staining was performed to verify the difference in LC3B expression among the DMSO, cirsiliol (20 µM), cirsiliol + SC79, and cirsiliol + siFOXO1 groups. Scale bars: 50 µm. The number of spots was counted on the fluorescent images using FIJI/Image J. Data are expressed as mean ± SD. *p < 0.05; **p < 0.01; ***p < 0.001