Fig 1: Topical Treatment With C1 and pan Eye Drops Increase the expression of TNFAIP3 and EP4. (A) Heatmaps showing differentially expressed genes between the vehicle (Veh) and Pan or C1 corneas. Gene analysis using the NanoString Mouse Inflammation panel v2 was performed on the cornea of mice subjected to desiccating stress for 5 days (n = 3–4/group). For all genes identified as significantly changed, log2 > 1.2 and padj. < 0.05. (B) Representative western blot in corneal epithelium lysates that were blotted with anti-TNFAIP3 or EP4 antibodies. Beta-actin was used for loading control. Densitometry of the three samples per group is shown in the bar graphs. Non-parametric Kruskal–Wallis followed by Dunn’s multiple comparison test. Each dot represents one biological sample containing cells pooled from four animals. P-value as shown. (C) Merged images of laser confocal microscopy of cornea cryosections stained with TNFAIP3 or EP4 antibodies (red) with DAPI nuclear staining (blue). Insets are high magnification to the adjacent area. Bar graphs on the right indicate fluorescence intensity normalized by the area of corneal epithelium. Non-parametric Kruskal–Wallis followed by Dunn’s multiple comparison test. Each dot represents one biological sample. P-value as shown. CJ, conjunctiva; Co, cornea; NC, negative control. (D) Merged images of laser confocal microscopy of whole-mount corneas stained with EP4 (red) and MHC II (green) antibodies with DAPI nuclear staining (blue). Z stacks were collected for the EP4 images and rotated 90 degrees to show spatial distribution. E, epithelium; S, stroma. (E) Merged images of laser confocal microscopy of the stroma of whole-mount corneas stained with EP4 (red) and CD209 (green) antibodies with DAPI nuclear staining (blue). For EP4 images, Z stacks were collected and images were rotated 90 degrees to show spatial distribution. E, epithelium; S, stroma.