Figure 1.

CXXC5 levels are inversely associated with the levels of β-catenin and wound-healing markers in human acute wounds. Wounded tissues (n = 5 samples) were excised from wound margins at 0, 7, 28, and 84 d after surgery in melanoma patients and subjected to H&E staining and immunohistochemical analyses. 0 d represents normal healthy biopsy (control). (A) Representative H&E and confocal (IHC) images (n = 5 independent experiments) for β-catenin, CXXC5, keratin 14, collagen I, or PCNA in the wounds are shown. Dashed lines indicate the epidermal–dermal boundary. F, fibroblasts; K, keratinocytes. (B and C, top) High magnification of representative IHC images for β-catenin, CXXC5, and the indicated wound-healing markers in keratinocytes (B) and fibroblasts (C) are shown. (bottom) Quantitative TissueFAXS analyses of IHC staining for β-catenin and CXXC5 (n = 5 independent experiments) were performed in five random, representative fields per each patient sample. Horizontal lines represent the mean fluorescence intensities. Student’s t test was used (***, P < 0.0005). (D and E, top) Representative images of IHC staining performed with antibodies against β-catenin (D) or CXXC5 (E) and markers for fibroblasts (vimentin), endothelial cells (CD34), or macrophages (CD68). Yellow arrowheads indicate the co-expression of β-catenin or CXXC5 and markers for fibroblasts, endothelial cells, or macrophages, and the green arrowhead indicates the cells expressing only CD68. (bottom) Quantitative TissueFAXS analyses of IHC staining for β-catenin and CXXC5 (n = 3 independent experiments) were performed in three random, representative fields (**, P < 0.005; ***, P < 0.0005). Means ± SD. Bars: (A–C) 100 µm; (D and E) 50 µm.