Pannexin1: Role as a Sensor to Injury Is Attenuated in Pretype 2 Corneal Diabetic Epithelium

Figure 4

Association of pannexin1 and P2X7 in unwounded corneas and 2 hrs after injury in DiO and B6 control corneal epithelium. Proximity ligation assays were performed using antibodies to P2X7 and pannexin1. (a) Representative enface and orthogonal (ortho) images of B6 control and DiO corneas are shown. The green color displays puncta of association. Corneal epithelium was counter stained with rhodamine phalloidin (asterisk marks wound area). The numbers and letters mark the individual cells that were analyzed using CellProfiler near the wound and back from the wound. The boxes indicate the representative individual cells that are shown in (b) after PLA analysis with CellProfiler. Bar equals 20 microns (PLA enface and actin) or 15 microns (PLA ortho). (b) Analysis of overlapping puncta for each condition (unwounded, leading edge, back from leading edge) was performed and a box plot drawn using R. The mean puncta for each region are placed above the box. are plotted, and two-way ANOVA with Tukey’s multiple comparison of means was performed. . is a minimum of 3 independent experiments.

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