Figure 7: Phosphorylation of ACF7 by FAK and Src plays an essential role in skin-wound healing in vivo.

(a) Wound healing as monitored by histological staining of skin sections at the wound edges 6 days after injury. Halves of wound sections are shown. Es, eschar. Dotted lines denote dermal–epidermal boundaries. Scale bar, 200 μm. (b) Quantification of the length of hyperproliferative epidermis generated at times indicated after wounding (upper panel). Error bars represent s.e.m. One-way ANOVA indicates that the difference between WT versus KO, or KO versus KO+ACF7 or KO+ACF7 versus KO+ACF7-YF is statistically significant (P<0.05). (c) Plotting of individual epidermal cell-movement trajectories in vivo. The arrow head indicates the direction towards wound centre. (d) Box and whisker plots of cell velocities in vivo. One-way ANOVA indicates that the difference between WT versus KO, or KO versus KO+ACF7 or KO+ACF7 versus KO+ACF7-YF is statistically significant (P<0.05). (e) Windrose plotting of cell-movement directions. The arrow head indicates the direction towards wound centre. (f) Quantification of cell-movement persistency in vivo. One-way ANOVA indicates that the difference between WT versus KO, or KO versus KO+ACF7 or KO+ACF7 versus KO+ACF7-YF is statistically significant (P<0.05). (g) Phase contrast images of skin-explant culture (day 4). Note outgrowth of epidermal cells is inhibited on loss of ACF7, which can be rescued by ACF7 but not by ACF7-Y259F mutant. Dashed lines denote boundary of skin explant (left) or leading edge of epidermal outgrowth from the explant (right). Scale bar, 50 μm. (h) Quantification of epidermal outgrowth in skin explants. Error bar represents s.d. One-way ANOVA indicates that the difference between WT versus KO, or KO versus KO+ACF7 or KO+ACF7 versus KO+ACF7-YF is statistically significant (P<0.05). ANOVA, analysis of variance.