Reduced CD44 on myeloid populations during early priming in the dLNs. (A) Gating strategy of MHCII^highCD11c^int migDCs, which were gated further for CD11b⁺ESAM^- migDC2b, CD11b⁺ESAM⁺ migDC2a, and CD11b-migDC1 in the dLNs 3 days after MOG_35-55 immunization. The mean percentage and SEM are shown in pink for each cluster (n = 4 per group). Doublets, dead cells, CD3+/CD19+ cells, eosinophils, and neutrophils were excluded. (B) Histogram and the MFI of CD44 expression on migDC1, migDC2a, and migDC2b. (C) Representative flow cytometry histograms showing CTV dilution of OT-II CD4⁺ T cells cocultured with resident DC2s (T + resDC2s) from dLNs from control or NIK^ΔCX3CR1 mice. Cells were pulsed with the indicated concentrations of OVA_323–339 peptide for 1 h prior to coculture. Division index of OT-II T cells, reflecting the average number of divisions per input cell. (D) Percentage of PD1+ OT-II cells from the antigen presentation assay. Data in B, C, and D are shown as the mean ± SEM and analyzed using two-way ANOVA with Šídák’s multiple comparisons test.  *P < 0.05, ***P < 0.001. Each dot represents one mouse. Data are from at least two independent experiments. MFI, median fluorescence intensity; CTV, CellTrace Violet.