In vivo chemotactic activity of mEAR2. Balb/c mice with well-formed air pouches were randomized into 3 groups (n = 3-5). The air pouches of mice were injected with 1 mL PBS alone or containing 1 μg/mL hANG or mEAR2, respectively. After 4 hours, the air pouches were washed with PBS containing 5 mM EDTA and 20 U/mL heparin. Cells recovered from the air pouches were counted and analyzed by FACScan after staining with PE–anti-CD11c and FITC–anti–I-A/I-E or isotypematched control antibodies. (A) Representative dot plots showing the PE and FITC double-positive DCs (top right quadrant highlighted by a rectangle) in the infiltrating cells recovered from PBS-, hANG-, or mEAR2-injected air pouches. (B) The average (mean ± SD) of total infiltrating cells or DCs of each group, which was calculated by multiplying total cell number with percentage of DCs of individual air pouches.