Figure 8. The synergistic anti-tumor effect of TRAIL and TSN in vivo.

(A) To construct xenograft model, A549 cells were inoculated into nude mice through subcutaneous injection. After the average tumor volume reached to 100 mm³, the animals were treated with 100 μg TRAIL, 0.173 mg/kg TSN, or both once per day for 24 consecutive days. Tumor volume (A) and animal weight (supplementary) were measured once per three days. Data represent the mean ± SEM from three independent experiments, **P < 0.01, as compared with mice treated with TSN alone. (B) The solid tumors removed from nude mice. (C) After treatment, tumor tissues were removed and lysed in RIPA lysis buffer. Western blot analysis was performed to evaluate the expression levels of the indicated proteins. Similar results were obtained in 3 independent experiments. (D) After treatments, tumor tissues were removed and subjected to Immunohistochemistry analysis. The representative image in 3 independent experiments was shown. (E) After treatment, the liver and spleen tissues were removed and subjected to H&E staining analysis. The representative image in 3 independent experiments was shown. (F) The different tissues removed from nude mice were dewaxed and hydrated, then stained with Hoechst and Annexin V-EGFP. The cell apoptosis was observed under fluorescence microscope. The representative image in 3 independent experiments was shown.