Z-GP-DAVLBH inhibits the AXL/AKT/GSK-3β/β-catenin pathway in osteosarcoma cells. (A)–(B) Osteosarcoma cells were treated with vehicle (0.1% DMSO) or Z-GP-DAVLBH (6 nmol/L) for 48 h. (A) The levels of p-AXL (Tyr779), AXL, p-AKT (Ser473), AKT, p-GSK-3β (Ser9), and GSK-3β in SJSA-1 and 143B cells treated with Z-GP-DAVLBH were determined by Western blotting analysis. (B) Representative blots of β-catenin, β-catenin (Ser552), and non-phosphorylated (active) β-catenin (Ser33/37/Thr41) in osteosarcoma cells. (C) and (D) Osteosarcoma cells after transfection with either NC siRNA or AXL siRNA were treated vehicle (0.1% DMSO) or Z-GP-DAVLBH (6 nmol/L) for 48 h. (C) Western blotting analysis was conducted to evaluate the effect of Z-GP-DAVLBH on the AXL/AKT/GSK-3β/β-catenin pathway components and (D) EMT-related markers in osteosarcoma cells. (E) and (F) Osteosarcoma cells were treated with Z-GP-DAVLBH (6 nmol/L) for 24 h. Transwell assays were conducted to evaluate the effect of Z-GP-DAVLBH on the (E) migration and (F) invasion capacities of SJSA-1 and 143B cells transfected with the indicated siRNAs. Scale bar: 200 μm. Data are presented as mean±SEM, n = 3; ∗P<0.05, ∗∗P<0.01, and ∗∗∗P<0.001 vs. the indicated groups.