Fig. 6: Effects of peptide-presenting cell on the development of CD8αα IEL precursors.

OT-I thymocytes depleted of mature CD8 single positives were stained with CPD450 and overlaid onto thymic slices from WT− → Kbm1, Kbm1 → WT, WT → WT, or Kbm1 → Kbm1 bone marrow chimeric mice. In some experiments, WT or Kbm1 mice were used instead of WT → WT or Kbm1 → Kbm1 control chimeras, respectively, with equivalent results. OVAp was added to thymic slices, which were harvested 48 h later for flow cytometric analysis. a Percent of OT-I thymocytes expressing CD122 and PD-1, displayed as values (left) or representative flow cytometry plots (right). b Expression of T-bet in OT-I thymocytes exposed to OVAp presented as Mean Fluoresence Intensity (MFI) of T-bet relative to no OVA controls (left), or representative flow cytometry plot (right). Dashed line indicates the average T-bet expression in no OVA control samples across all treatment conditions c Proliferation of OT-I thymocytes displayed as percent of OT-I thymocytes that have undergone 3 cellular divisions, determined based on dilution of CPD450 according to the gate shown in the representative flow cytometry plot (right). Data are pooled from (left panels) or representative of (right panels) 3 (WT → Kbm1 and Kbm1 → WT conditions) or 5 (no OVA, WT → WT, Kbm1 → Kbm1 conditions) experiments with mean and SEM of n = 12–22 total slices, where each dot represents an individual slice. ns not significant (p > 0.05), *p < 0.05, ***p < 0.001, ****p < 0.0001 (one-way ANOVA with Bonferroni’s correction).