Fig. 6. Cultures and analyses of primary keratinocytes. (A) In vitro scratch assay. Both the wild-type and mutant keratinocytes migrated into the gap within 20 hours. (B-E) Laser scanning microscopic analysis of actin organization in wild-type and mutant keratinocytes at the leading edge of a scratch assay performed as above 5 hours post-scratch. The cells were fixed and stained with phalloidin. Lamellipodia (B,C) and filopodia (D,E) are observed in both the wild-type and mutant keratinocytes, although in the mutant the lamellipodia are often not oriented toward the gap (arrows in C; arrowheads in B for comparison) and the filopodia appear thinner. Scale bars: 100 µm (A; all images are the same magnification); 50 µm (B,C); 10 µm (D,E).