Flow cytometry analysis to determine the number of E7-specific CD8+ T cells in the splenocytes of mice vaccinated subcutaneously or intratumorally. The TC-1 tumor-bearing C57BL/6 mice (5 per group) were immunized subcutaneously or intratumorally using a combination of 20 μg/mouse of HPV-16 E7 (aa 49-57) peptide and 20 μg/mouse of PADRE peptide with 20 μg/mouse of poly(I:C) twice with a 1-week interval. One week after the last vaccination, splenocytes were harvested and characterized for E7-specific CD8+ T cells using intracellular IFN-γ staining followed by flow cytometry analysis. (A) Representative flow cytometry data showing the number of E7-specific IFNγ+ CD8+ T cells in splenocytes from mice vaccinated subcutaneously or intratumorally (right upper quadrant). (B) Bar graph depicting the numbers of E7-specific IFN-γ-secreting CD8+ T cells per 3 × 104 pooled splenocytes (mean ± s.d.). * indicates p < 0.05. Data shown are representative of two experiments performed.