Therapeutic effects of TSP-2 shRNA in the MBT-2 mouse tumor model. (a) TSP-2 shRNA abolished the expression of TSP-2 in vitro. HA-TSP-2 expression was detected by immunoblotting. (b) Tumor-bearing mice were treated with the indicated shRNA, and the tumor sizes (*P < 0.05, TSP-1 shRNA versus TSP-2 shRNA) and (c) survival rates (*P < 0.05, TSP-1 shRNA versus TSP-2 shRNA) were determined. (d) TSP-1 shRNA induces stronger CTL response than TSP-2 shRNA. Effector cells were splenocytes isolated from mice with the indicated treatments. The target cells were MBT-2 luciferase cells (**P < 0.01, control versus TSP-1 shRNA in the ratio 25:1; **P < 0.01, TSP-1 shRNA versus TSP-2 shRNA in the ratio 50:1, n = 3). (e) Analysis of regulatory CD4+ Foxp3+ T cells in mice that received TSP-1 or TSP-2 shRNA. Left panel: representative flow cytometry analysis of the percentage of CD4+ Foxp3+ T cells in the lymph nodes. Right panel: the bar graph represents average + SEM of the percentage of CD4+ Foxp3+ T cells (*P < 0.05; n = 3 mice per group). NS, no statistical difference; Scr, scramble; shRNA, short hairpin RNA; TSP-1, thrombospondin 1.