FIGURE 6:
Reduced contribution of ILK-deficient cells to the regenerated epidermis. The dorsal skin of P50 K15.CrePR1-YFP-Ilkf/+ or K15.CrePR1-YFP-Ilkf/f mice was treated daily with topical RU486 for 5 d. Five days after the last treatment, the mice received a full-thickness excisional wound with a 6-mm biopsy punch. (A) Tissues were harvested 9 d postwounding and processed for immunofluorescence microscopy using antibodies against Ki67. YFP was detected using anti-GFP antibodies. Arrows indicate wound margins. Continuous and dashed lines indicate, respectively, the cornified layer and the dermal–epidermal junction. The box in the panels on the left represents regions shown at higher magnification in the adjacent panels at right. (B) Tissues were harvested 12 d postwounding and processed for immunofluorescence microscopy using anti-GFP antibodies to visualize YFP. The arrow in the micrographs indicates areas shown below at higher magnification. Solid and dashed lines represent the outer surface of the epidermis and the dermal–epidermal junction, respectively. Nuclear DNA was visualized with Hoescht 33342. Bars, 100 μm. (C) The number of YFP-positive cells in the reepithelialized epidermis between the wound margins of mice with the indicated genotype was scored. The results are expressed as the percentage of YFP-expressing cells (basal plus suprabasal) relative to total cell number (basal plus suprabasal). Total cell numbers were assessed by counting the Hoescht 33342–stained cell nuclei. The data are shown as the mean plus SEM (n = 8).