Fig. 2: Ezh2 helps establish memory properties in activated CD8+ T cells early during expansion | Nature Communications

Fig. 2: Ezh2 helps establish memory properties in activated CD8+ T cells early during expansion

From: Ezh2 phosphorylation state determines its capacity to maintain CD8+ T memory precursors for antitumor immunity

Fig. 2

. a Schematic diagram of three characteristic phases of the T-cell response and the possible role of Ezh2 in each phase. b–g WT and Ezh2 −/− naive Pmel-1 cells (Thy1.1+) were transferred into sublethally irradiated non-tumor-bearing B6 mice (Thy1.2+), followed by immediate treatment with IL-2 and gp100-DCs for 3 days. Donor T cells were recovered at 4 days and 7 days after transfer. Plots and graphs show the percentage of KLRG-1-expressing (KLRG1hi) TCMP and TEFF in the spleen (b) and PB (c). d Graphs show the percentage and numbers of TCMP and TEFF (left panel) in the spleen at 4 days and 7 days after transfer. The right panel shows the percentage and numbers of MPCs and SLECs measured with KLRG-1 and CD127 at 4 days and 7 days after transfer. e The percentage of Annexin V-positive cells in the subpopulation of TCMP and TEFF at 4 days and 7 days after transfer. f Real-time RT-PCR measurement of p19 Arf in the subset of TCMP and TEFF of 4 days and 7 days. g Histograms show the expression of indicated surface markers on WT and Ezh2 −/− T cells derived from the spleen at 4d after transfer. h, i WT and Ezh2 −/− naive Pmel-1 cells (Thy1.1+) were transferred into sublethally irradiated non-tumor-bearing B6 mice (Thy1.2+), followed by immediate treatment with IL-2 and gp100-DCs for 3 days. By 7 days after transfer, donor TCMP and TEFF were highly purified using FACS sorter, and transferred into sublethally irradiated secondary recipients that had been immunized with gp100-DCs 7 days earlier (described in Supplementary Fig.4). Forty-two days later, donor T cells were collected from the spleen of these secondary recipients (h), and further cultured ex vivo for additional 5 days (i). Plots and graphs show the frequency of donor T cells derived from the secondary recipients of WT and Ezh2 −/− TCMP and TEFF. *p < 0.05, **p < 0.01, and ***p < 0.01 (two-tailed unpaired t test). The data are representative of four independent experiments with n = 3 mice per group in each (b–g; mean ± SD) or two experiments with n = 4 mice per group in each (h, i, mean ± SD)

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