AMD3100 enhances the antitumor effect of doxorubicin in an orthotopic OS mouse model. (A) Macroscopic appearance of OS tumors in the tibia of C3H mice after 8 consecutive injections of PBS, 5 mg/kg AMD3100, 1 mg/kg doxorubicin and AMD3100 plus doxorubicin. (B) The body weight of the mice in each group was measured at every injection, and the growth curve was obtained. (C) The volume and weight of the tumors in the four treatment groups were determined. (D) The expression levels of C-X-C motif chemokine receptor 4, beclin 1, light chain 3B and P-glycoprotein in tumor tissues were detected by immunohistochemical staining, and the areal density was quantified and subjected to statistical analysis. (E) The expression of p-PI3K, p-AKT and p-mTOR in tumor tissues was detected by immunohistochemical staining, and the areal density was quantified and subjected to statistical analysis. *P<0.05, **P<0.01, ****P<0.0001. OS, osteosarcoma; p-, phosphorylated.