Fig. 8.

CCL chemokine expression levels are altered in wounded Mmp12^−/− corneas. (A) Relative expression levels of CCL chemokine family members CCL2, CCL7, CCL8 and CCL20 in WT and Mmp12^−/− corneas before injury (d0) and 6 days after injury (d6), as determined by qRT-PCR. Expression levels are relative to uninjured WT corneas. Results are means ± s.e.m. (n = 6), **P<0.005 and *P<0.05. (B) Cytokine expression in WT and Mmp12^−/− corneas 1 day after injury. Individual cytokines are spotted in duplicate and the identity of CCL2 is indicated. Positive control spots are located at the corners. (C) Relative cytokine levels in corneas of WT and Mmp12^−/− mice 4 days after injury. Duplicate spots were averaged. CCL2 levels are indicated with an asterisk (*). (D) Effect of CCL2 neutralization on macrophage infiltration in the cornea. An antibody to CCL2 or PBS control was injected in the subconjunctival space of WT mice. After 2 hours, the corneas were wounded chemically. The subconjunctiva were re-injected with an antibody to CCL2 or PBS control 3 days after injury and the corneas were collected 6 days after injury. Representative whole-mount micrographs of the corneas demonstrate decreased F4/80+ cells following CCL2 antibody injection compared with the PBS control. (E) Quantification of F4/80 levels in corneas of PBS-treated and anti-CCL2-treated mice. The mean number (± s.e.m.) of F4/80+ cells per corneal section are shown and are 128±38 pixels (PBS; n = 7) and 28±8 (anti-CCL2; n = 7), *P<0.05. Scale bars: 10 µm.