Intracellular cytokine staining followed by flow cytometry analysis for TERT198-specific CD8+ T cell immune responses. 5-8 weeks old C57BL/6 mice were vaccinated with 5 μg of L1 protein of HPV16-CRT/TERT198, or HPV16-CRT pseudovirions via footpad injection, and boosted twice at 4-day interval with the same regimen. 1 week after last vaccination, splenocytes were prepared and stimulated with TERT198 peptide (1 μg/ml) at the presence of GolgiPlug overnight at 37 C. The TERT198-specific CD8+ T cells were then analyzed by staining surface CD8 and intracellular IFN-γ. A. Representative flow cytometry data. B. Graphical representation of the number of TERT198-specific CD8+ T cells per 3 × 105 splenocytes. Data expressed as means ± standard deviations (SD) are representative of at least two different experiments.