Immune priming in the draining LNs is reduced in CCX-CKR^−/− mice. (A) Viable cell counts from inguinal LNs during the time course of EAE. Six mice per time point per group were analyzed. Data shown are the mean cell number ± SEM (**P < .01). (B) Six mice per group were immunized for EAE and fed BrdU at 0.8 mg/mL in their drinking water from day 6. On day 9, draining LNs were harvested, stained for CD4⁺ and BrdU, and analyzed by flow cytometry. Representative dot plots are shown. Data shown are the mean proportion of CD4⁺/BrdU⁺ cells ± SEM (**P < .01). (C) Ex vivo proliferation assay to MOG_35-55 using cells from mice taken on days 5 and 28 postimmunization for EAE. Cells taken from 6 immunized mice per time point per group were analyzed. Data shown are the mean percentage of CD4⁺ cells that proliferated during the 4-day incubation ± the SEM (* P < .05). (D) On day 12 postimmunization for EAE, draining LNs were harvested, viable cells counted, and cells cultured overnight at 4 × 10⁶ /mL in complete RPMI at 37°C, 5% CO₂. Cells were then harvested and stained for flow cytometric analyses. Data shown are from 11 mice per group in total from 2 independent experiments pooled and represent the mean ± SEM (**P < .01). Representative dot plots showing the gating for CD4 and CCR6 are shown.