Supplementary Figure 3: AP4 is essential for the secondary population expansion of memory CD8+ T cells.
From: c-Myc-induced transcription factor AP4 is required for host protection mediated by CD8+ T cells
(a) Flow cytometry analysis of splenocytes showing expression of CD8α, CD127, CD62L and H-2Db–gp(33–41)-specific TCR on day 60 after LCMV-Arm infection. (n = 4). (b,c) Statistical analysis of absolute numbers of gp(33–41)-specific total memory CD8+ T cells and CD62L+ central memory CD8+ T cells in the spleen on day 60 after LCMV-Arm infection (b) and frequencies of gp(33–41)-specific memory CD8+ T cells in peripheral blood mononuclear cells (PBMC) in Tfap4–/– (n = 6), Tfap4F/F CD8-Cre+ (n = 7) or control C57BL/6 mice (n = 14) on day 60 after LCMV-Arm infection (c). (d) IFN-γ ELISPOT assay showing frequencies of OVA-specific memory CD8+ T cells in splenocytes of Tfap4–/– (n = 4) and control C57BL/6 mice (n = 4) on day 45 after LM-OVA infection. All mice had less than 11 spots per million splenocytes when no antigen was included. (e,f) Flow cytometry analysis showing expression of CD8α, CD45.2, CD62L, KLRG1 and H-2Db–gp(33–41)-specific TCR in splenocytes of CD45.1 wild-type host mice on day 5 after LCMV-Arm infection. Before infection, the host mice received CD8+ T cells from Tfap4–/– or C57BL/6 mice, which were infected with LCMV-Arm 60 days before. (n = 8 in two independent experiments). Statistical analysis is shown in (f). (g) Flow cytometry analysis of PBMC of Tfap4–/– (n = 15), Tfap4F/F CD8-Cre+ (n = 9), or control C57BL/6 mice (n = 14) 5 days after secondary challenge with LM-OVA. (h) Statistical analysis of numbers of total CD8+ T cell and OVA-specific CD8+ T cells numbers in blood and spleen 5 days after secondary challenge with LM-OVA is shown. Each symbol represents an individual mouse; small horizontal lines indicate the mean (± s.d.). * P < 0.01, ** P < 0.001, *** P < 0.0001, NS: not significant by unpaired t-test (b,d,f) or one-way ANOVA (c,h). Data are representative of three experiments.
