Figure 5

Prediction and validation of DEDD as target gene of miRNA-15b. A) Schematic representation of miRNA-15b’s sequence bound to DEDD. B) The luciferase constructs fused to 3^′UTR (or mut) of DEDD were co-transfected in 293 T cells with mimic-15b or mimic-NC. The luciferase activities were expressed as mean ± standard deviation (n = 3) of the ratio of firefly luciferase over renilla luciferase (*P < 0.01). C) CD8+ T cells were isolated from C57BL/6 mice and transfected with mimic-NC or mimic-15b, the levels of DEDD mRNA in T cells with mimic-NC or mimic-15b were measured by qRT-PCR or PCR electrophoresis (72 h after transfection, * P < 0.05). D) The levels of DEDD protein in CD8+ T cells with mimic-15b or mimic-NC were assessed by western blot (72 h after transfection, * P < 0.01). E) CD8+ Te and Tm were prepared from the spleens of C57BL/6 mice with Lewis lung carcinoma. The levels of DEDD protein in CD8+ Te or Tm were assessed by western blot (* P < 0.05).