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. 2016 Jun;186(6):1466–1480. doi: 10.1016/j.ajpath.2016.01.019

Figure 2.

Figure 2

Altering IL-1 signaling in normal healing cornea affects corneal epithelial wound healing rate and gene expression in corneal epithelial cells. A: Normal (NL) mice were subconjunctivally injected with 5 μL IgG (200 ng/μL), left eyes, and IL-1β neutralizing antibody (200 ng/μL), right eyes, once at −4 hours, or injected with non-specific control siRNA (20 μmol/L), left eyes, and IL-1 receptor antagonist (IL-1Ra)–specific siRNA (20 μmol/L), right eyes, twice at −24 and then −4 hours. Epithelial samples for each condition were collected. Representative images of wounds at 20 hours after wounding (hpw). B: Remaining wounds were calculated and analyzed with paired t-test. C: Enzyme-linked immunosorbent assay confirmation of siRNA knockdown of IL-1Ra (all isoforms) on the protein level, analyzed with one-way analysis of variance. Expressions of IL-1β (D), IL-1Ra-V1 (E), and IL-1Ra-V2 (F) in naïve (NL), IgG control (IgG), mouse IL-1β neutralizing antibody (α-Iβ) treated, control, non-specific siRNA (CTsiR), and IL-1Ra siRNA (si1Ra) treated corneal epithelial cells (CECs) assessed by real-time quantitative PCR and analyzed with one-way analysis of variance. Data are given as means ± SEM (B and D–F). n = 5 (B); n = 3 (D–F). ∗P < 0.05, ∗∗P < 0.01, and ∗∗∗P < 0.001.