Fig. 4

Immunolocalization of macrophages and analysis of keratocyte phenotype in healed cornea at 30 hours post-debridement. Immunohistochemistry detects F4/80-labeled macrophages, aldehyde dehydrogenase 3A1 (ALDH3A1, keratocyte marker), and -smooth muscle actin (SMA, myofibroblast marker). F4/80-labeled macrophages were more frequently observed beneath the regenerated epithelium in the KO mouse (b), while few F4/80-labeled cells were seen in the WT corneas (a). The cells in the posterior stroma were labeled for ALDH3A1, while fibroblastic cells in the anterior stroma were not labeled in the WT cornea (c). In the KO cornea, the majority of the cells in the stroma were negative for ALDH3A1 (d). A few -smooth muscle actin-positive myofibroblasts were detected in the WT stroma (e). Almost all the stromal cells were labeled with anti-SMA antibody and thus were myofibroblasts in the KO cornea (f). Bar, 50 μm. Reproduced from Shirai et al. [9]