The number of LECs increases in inflamed ears of VEGF-A Tg mice. A DTH response to oxazolone was induced in the ears of VEGF-A Tg mice. Ear tissue was analyzed 9 days after challenge. (A) At this time point, the inflamed ears were markedly increased in thickness compared with ears from control (ctr) animals. (B) FACS analysis was performed on cells derived from enzymatically digested ears (example shown: inflamed ears). Cells were stained for CD45, CD31, and podoplanin to differentiate between leukocytes (CD45⁺CD31⁻podoplanin⁻), BECs (CD45⁺CD31⁺podoplanin⁻), and LECs (CD45⁺CD31⁺podoplanin⁺). (C-E) Quantitative FACS analysis revealed that total numbers of leukocytes (C), BECs (D), and LECs (E) were increased in cell suspensions of inflamed ears compared with those of control ears. *P < .05; ***P < .001 (compared with control). (F) Immunofluorescence analysis of LYVE-1 (green) and MECA-32 (red) expression confirmed that vascularization was increased in inflamed compared with control ears. Scale bars represent 50 μm. Error bars are SE.