Role of early Notch signaling in CD8⁺ T cells during Lm-OVA infection. (A) Staining of the Notch2 extracellular domain in Notch1/2 WT CD8⁺ OT-I T cells on day 2 after infection with Lm-OVA. (B) Staining of the Notch2 transmembrane and intracellular domain in Notch1/2 WT and KO CD8⁺ OT-I T cells on day 2 after infection with Lm-OVA. (C–E) Proportion of SLECs (C), proportion of CX3CR1^hiCXCR3^lo SLECs (D), and MFI of 1B11 (E) from mice infected with Lm-OVA and treated with anti-DLL1/4 blocking antibodies on days (d) 0 and 2, day 2, day 3, or day 3.5. Untreated Notch1/2 WT and KO mice were used as control. (F) Representative histograms (left) and compilation (right) of the proportion of KLRG1⁺ cells among effector OT-I CD8⁺ T cells on day 3 after Lm-OVA infection. (G) Differential transcription of classical Notch target genes in the presence or absence of Notch signaling in day 3 effector CD8⁺ T cells. (H) Proportion and numbers of Notch1/2 WT and KO OT-I CD8⁺ T cells recovered on day 3 after infection with Lm-OVA. (I) Proportion of Notch1/2-deficient or sufficient OT-I CD8⁺ T cells positive for 7-AAD on day 3 after infection with Lm-OVA. Data are from one (G and J), two (C–F and I), or three (H) independent experiments with a total of n = 2–8 mice per group. Error bars display means ± SEM. Ordinary one-way ANOVA with Tukey’s multiple comparisons was used for multiple group comparison and unpaired two-tailed t test was used for two-group comparisons. ***P < 0.001, ****P < 0.0001.