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Fig. 4. HB-EGF contributes to the epithelial sheet migration in leading edge formation. (A) Schematic diagram showing how the progression of eyelid formation was measured. Contribution of leading edge formation and root formation to total eyelid formation was estimated as shown. (B) Progression of total eyelid formation (left panel), leading edge formation (middle panel) and root formation (right panel) in HB^del/+ (blue dots, n=19) and HB^del/del (orange dots, n=20) animals. Horizontal bar in the graph indicates the mean value. (C) HB-EGF promotes F-actin reorganization in the developing eyelid epithelium. Whole-mount staining of HB^del/+ (a-d) and HB^del/del (e-h) embryos at E15.0 (a,c,e,g) and E15.5 (b,d,f,h) was performed using FITC-phalloidin and propidium iodide to visualize F-actin and DNA, respectively. High-magnification views of boxed areas in a,b,e,f are shown in c,d,g,h, respectively. Both actin cable formation (white arrowheads) and radial F-actin fiber (black arrowheads) can be detected more clearly in HB^del/+ eyelids than in HB^del/del eyelids. Scale bars: 500 µm for a,b,e,f; 200 µm for c,d,g,h.

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