T-bet and ZEB2-mediated control of S1PR5 expression in CD8⁺ T cells. (A) Mice were adoptively transferred with naive GFP⁺ OT-I (OT-I WT) and CD45.1⁺ OT-I Tbx21^−/− (OT-I Tbx21^−/−) and infected with HSV-OVA. Expression of indicated genes quantified by qPCR on OT-I T cells from the spleen 8 dpi. (B) Effector gBT-I T cells were transduced with Ctrl-RV or Tbet-RV GFP-expressing RVs and cotransferred into mice infected with HSV. Expression of indicated genes quantified by qPCR on sort-purified gBT-I T cells from the spleen 8 dpi. (C) Effector gBT-I T cells were nucleofected with control-nontargeting (CD45.1⁺CD45.2⁺ gBT-I sgCtrl) or Zeb2-targeting (CD45.1⁺ gBT-I sgZeb2) sgRNA/Cas9 RNPs and cotransferred into HSV-infected mice. Expression of indicated genes quantified by qPCR on sort-purified transgenic T cells from the spleen 8 dpi. (D) Effector gBT-I T cells were transduced with control or ZEB2 (ZEB2-RV) RVs and maintained in culture with IL-15 for 3 d. Expression of indicated genes was quantified by qPCR in GFP⁺ gBT-I T cells. (E) Effector gBT-I T cells were transduced with control or Tbet-RV and nucleofected with control-nontargeting (CD45.1⁺CD45.2⁺ gBT-I sgCtrl) or Zeb2-targeting (CD45.1⁺ gBT-I sgZeb2) sgRNA/Cas9 RNPs and cotransferred into HSV-infected mice. S1pr5 expression was quantified by qPCR in sort-purified gBT-I T cells from the spleen 8 d after transfer. In A–C and E, data are representative of two independent experiments, with n = 4–5 mice per experiment. In D, data are pooled from two to three independent experiments, with n = 2–3 samples per condition. *, P < 0.05, **, P < 0.01 by Mann–Whitney test. A.U., arbitrary units; rel., relative.