Fig. 3
CD11bhiLy6Chi cell depletion augments corneal nerve loss while increasing IL-6. A, B Representative photographs of corneas before and after lissamine green vital dye staining (A). The green-stained areas indicate corneal epithelial defect. Quantification of corneal opacity at 7 d post-injury as graded by the standardized scoring system (0–4) (B). C, D Representative β-tubulin III and Ly6G staining images of corneal whole-mounts, with corneal center (blue box) and periphery oriented to limbus at top (red box) at 7 d post-injury (C). Analysis of corneal innervation and Ly6G+ cell infiltrates at 7 d post-injury quantified as percentage threshold area positive for β-tubulin III and Ly6G, respectively (D). E, F Representative flow cytometry cytograms (E) and quantitation of CD11bhiLy6Ghi cells and CD11bhiLy6ChiLy6Glo cells in peripheral blood at 1 and 7 d post-injury (F). G, H Real-time RT-PCR for pro-inflammatory chemokine/cytokine mRNAs in cornea and blood cells at 1 d post-injury. The mRNA levels are presented as fold changes relative to the levels in normal corneas or blood cells without injury. Mean values ± SD are presented, where each circle represents the data from an individual mouse. A white circle depicts the data from an uninjured, naive mouse, a blue circle from an injured mouse treated with isotype control IgG (2A3), and a red circle from an injured mouse treated with anti-Ly6C Ab (Monts-1). *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001, ns: not significant, as analyzed by one-way ANOVA and Tukey’s multiple-comparison test