The same murine samples shown in Fig. 3 were tested by multicolor ICS assay for production of INF-γ, TNF-α and CD107a. (A) or (C) are shown for each individual combination of functions of HIV clade C Gag-specific CD8 T cellsin PBMCs or splenocytes, respectively. (B) or (D) are shown for each combination of functions (three, INF-γ+TNF-α+CD107a+; two, INF-γ+TNF-α+,INF-γ+CD107a+ and TNF-α+ CD107a+; one, INF-γ+, TNF-α+and CD107a+) of HIV clade C Gag-specific CD8 T cells in PBMCs or splenocytes, respectively. These were measured by multicolor ICS assay following stimulation with clade C p24 peptide. IFN-γ, TNF-α and CD107a were used as the functional markers. This assay was performed in 5-6 individual mice. Data represent the means ± S.E.M. and are representative of two independent experiments. *, p<0.05; **, p<0.01.