CD8+ T-cells were isolated from control and miR-17-92/Pmel-Tg mice. (A) Frequencies of gp100 tetramer+ cells were analyzed by flow cytometry. Flow data are representative of three independent experiments. (B) Expression levels of miR-17-5p were measured by qRT-PCR. (n=3, ***P < 0.001, t-test) (C) CD8+ T-cells were stimulated with hgp10025-33 (0.125 μg/ml) in the presence of feeder cells for 46 hours and evaluated for IFN-γ production by ELISA. (*P < 0.05, t-test) (D) Antigen-specific cytotoxicity of CD8+ T-cells without in vitro stimulation was evaluated against GL261 glioma cells loaded with or without hgp10025-33 peptide by a 6-h 51Cr-release assay. (**P < 0.01, t-test) (E) CD8+ T-cells were cultured with IL-2 (100 IU/ml) and hgp10025-33 peptide (0.125 μg/ml) in the presence of feeder cells. After 6 days, these cells were harvested and evaluated for antigen-specific cytotoxic activities. (*P < 0.05, t-test) Graphs are presented as mean+/−SD from at least two independent experiments.