Flow cytometry analysis to determine the number of E7-specific CD8+ T cells in the tumor-infiltrating lymphocytes 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, TILs were harvested and characterized for E7-specific CD8+ T cells using intracellular IFN-γ staining followed by flow cytometry analysis. (A) Bar graph depicting the numbers of tumor-infiltrating CD8+ T cells from mice vaccinated either subcutaneously or intratumorally (mean ± s.d.). (B) Representative flow cytometry data showing the percentage of E7-specific IFNγ+ CD8+ T cells in tumor-infiltrating lymphocytes derived from mice vaccinated subcutaneously or intratumorally (right upper quadrant). (C) Bar graph depicting the numbers of tumor-infiltrating E7-specific CD8+ T cells from mice vaccinated either subcutaneously or intratumorally (mean ± s.d.). Data shown are representative of two experiments performed. * indicates p < 0.05.