Fig. 4
IL-6 neutralization prevents corneal nerve damage and IL-6 replenishment exacerbates it. A, B Representative corneal photographs without and with lissamine green staining for visualization of epithelial defect at 7 d post-injury (A). Severity of corneal opacity and epithelial defect as graded by standardized scoring systems (0–4 for corneal opacity and 0–3 for epithelial defect) (B). C–F Representative images of corneal whole-mounts, center (blue box) and periphery (red box) after β-tubulin III (C), IL-6 or Ly6G staining (E). The tissues were extracted at 7 d post-injury. Quantitative analysis of corneal nerve density (D) and corneal infiltration of Ly6G+ cells and Ly6C+ cells (F). G, H Flow cytometric analysis for CD11bhiLy6Ghi cells and CD11bhiLy6ChiLy6Glo cells in blood at 7 d post-injury. Mean values ± SD are shown, and each circle depicts the data from an individual mouse. A white circle indicates the data from control mice (either naive mice without injury or injured mice receiving isotype control IgG). A red circle indicates the data from another control mice receiving HBSS (Hank’s balanced salt solution, vehicle for recombinant mouse (rm) IL-6) after injury. A blue circle demonstrates the data from a mouse receiving anti-IL-6 Ab, and a green circle from a mouse receiving rmIL-6. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001, ns: not significant, as analyzed by one-way ANOVA and Tukey’s test or Mann–Whitney U test