Fig 2: MMP13 promotes LSCs proliferation and migration. (A) Pie chart showing 192 genes of group III are annotated as interferon regulated genes. (B) Top 10 interferon regulated genes preferentially upregulate in wounded cornea. The size of the dot represents the gene expression. (C) Representative immunofluorescence staining of P63, PAX6, and Ki67 expression in LSCs. Scale bar, 50 µm. (D) The qPCR analysis of top 10 genes expression in recombinant IFNß treated LSCs (n = 3). Data are represented as mean ± SE. (E) Western blot analysis of MMP13 expression in recombinant IFNß treated LSCs at indicated time points. (F) FACS analysis of carboxyfluorescein succinimidyl ester (CFSE) labeled LSCs. The original fluorescence intensity at d0 represents the starting point. The control group and recombinant MMP13 treated group are analyzed at d3. Representative data from three independent experiments are shown. (G) Scratch wound assays of control LSCs and recombinant MMP13 treated LSCs. Representative images of LSCs at indicated time-points. Scale bar, 500 µm. (H) Quantifications of wound closure of LSCs are shown (Students t-test, n = 3). Data are represented as mean ± SE (*P < 0.05).

Fig 3: MMP13 enhances corneal wound closure. (A) The qPCR analysis of Mmp13 in unwounded and wounded cornea at d1 post-injury. (B) Representative immunofluorescence staining of MMP13 and KRT6 in unwounded and wounded cornea at d1 post-injury. Scale bar, 100 µm. (C) Western blot analysis of MMP13 proteins in unwounded cornea and wounded cornea treated with or without IFNß antibody. Representative immunoblots are presented, n = 3. (D) Representative images of the epithelial defect in wounded corneas of control, MMP13 antibody, IFNß antibody and IFNß antibody combined with recombinant MMP13 treatment (for each treatment: upper panel, white light micrograph; lower panel, fluorescein staining of corneal surface). (E) Quantification of re-epithelialization in control, MMP13 antibody, IFNß antibody, and IFNß antibody combined with recombinant MMP13 treated mice corneas. Epithelial defect is presented as the percentage of the original wound size (Students t-test, n = 5). Data are represented as mean ± SE (*P < 0.05, **P < 0.01, ***P < 0.001).