Inflammation-induced up-regulation of CCL21 protein does not correlate with DC migration. (A) Ear whole-mount immunofluorescence revealed abundant expression of CCL21 in lymphatic vessels in control and CHS- or CFA-inflamed ears. Scale bar represents 50 μm. (B) Quantification of CCL21 protein levels in ear lysates generated from uninflamed, CHS-inflamed, or CFA-inflamed ears of mice (n = 3 mice per group). Tissue inflammation led to a 2.3-fold (CHS) and 1.7-fold (CFA) increase in CCL21 protein. (C-D) FITC painting experiments were performed in uninflamed and CHS-inflamed or CFA-inflamed ears of mice. (C) LN single-cell suspensions were analyzed by FACS for the presence of I-A/I-E⁺CD11c⁺FITC⁺ cells. The number within each plot represents the percentage of gated cells. (D) Quantification of the total numbers of I-A/I-E⁺CD11c⁺FITC⁺ DCs detected stronger DC migration in response to CFA- compared with CHS-induced tissue inflammation. (E) CD11c⁺ cells in CHS or CFA-inflamed ears and control ears were analyzed by quantitative FACS analysis of ear single-cell suspensions. *P < .05. ***P < .001.