Fig. 3. Although ß4 integrin localization appears normal during migration, subtle differences in the localization of 9 integrin emerge between the wild-type and synd1ko corneas immediately prior to wound closure. (A) Unwounded tissues and tissues taken at 18 hours after wounding are double-labeled to detect synd1 and ß4 integrin simultaneously. Asterisks indicate the regions shown at higher magnification at the right — both as doubly and individually labeled images to allow for better determination of the regions of overlap. Arrows indicate the tip of the leading edge; migration occurs from left to right. Bar, 80 µm in lower magnification images at the left and 25 µm in higher magnification images at the right. (B) En face views of whole mounts of corneas from control or wounded mice have been treated with propidium iodide to highlight nuclei and simultaneously stained with an antibody to detect 9 integrin. 9 integrin is absent from the central corneas of unwounded wild-type animals but present at the limbal region as seen in the inset. Similar results are seen in the synd1ko mouse. When matched for the size of the remaining wound, wild-type corneas show more 9 integrin around epithelial cells close to the leading edge (asterisks) as well as at sites where seams form as advancing epithelial sheet edges merge (arrows) compared to the synd1ko corneas. Trapping of sera at the leading edge within wound opening occurs in some samples and is not specific. Bar, 150 µm.

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