Characterization of the E7-specific CD8+ T cell immune responses following vaccination with CRT/E7 DNA with or without imiquimod in tumor-bearing mice. C57BL/6 mice were divided into four groups (5/group: untreated, imiquimod alone, CRT/E7 alone, and combined group). All mice were inoculated with TC-1 cells (1 × 104/mouse) s.c. over right leg at d0. Treatment schedules were the same as in Figure 1. Splenocytes from tumor-bearing mice were harvested at d24 for immune response analysis, stimulated with E7 epitope (aa49-57), and stained for both CD8 and intracellular interferon-γ (IFN-γ). A) Representative flow cytometry data demonstrating the percentage of CD8+IFN-γ+ T cells. The numbers of CD8+ IFN-γ+ double positive T cells are indicated in the upper right hand corner of each representative dot plot. B) Bar graph representing the number of E7-specific CD8+ T cells/3 × 105 splenocytes (*, p < 0.05). Representative data from one of three independent experiments are shown.