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. 2015 Jun 29;212(7):1061–1080. doi: 10.1084/jem.20141601

Figure 9.

Figure 9.

Co-treatment with PTD-DBM and VPA accelerates cutaneous wound healing and synergistically induces collagen production in mice. The wounded skin of 7-wk-old male C3H mice was treated daily with 100 µM PTD-DBM and/or 500 mM VPA or with 100 µM EGF for 11 d (n = 16 mice/group). (A) IHC staining of wounds treated with PTD-DBM was performed to detect β-catenin. Representative images are shown (n = 3 independent experiments). (B) Representative H&E-stained images of wounds at 12 d after wounding with the different treatments are shown (n = 3 independent experiments). The arrowheads represent the edges of the wounds. (C) Representative images of macroscopic wounds and H&E and IHC staining (n = 5 mice group) for β-catenin, keratin 14, collagen I, PCNA, or p-Erk in wounds 12 d after wounding are shown. Dashed lines demarcate the epidermal–dermal boundary (n = 3 independent experiments). F, fibroblasts; K, keratinocytes. (D) Relative wound closure rates for wounds from mice treated with PTD-DBM and/or VPA or EGF were quantified as percent wound closure as shown. Wound sizes were measured at 1, 3, 5, 7, 9, and 12 d after wounding (*, P < 0.05; **, P < 0.005; ***, P < 0.0005; n = 16 mice/group). (E) Western blot analyses of β-catenin, keratin 14, α-SMA, collagen I, PCNA, endothelin-1, c-Myc, cyclin D1, p-Erk, and Erk in wounds (n = 2 mice/group) 12 d after wounding were performed (n = 2 independent experiments). Relative densitometric ratios of each protein to Erk protein are shown. (F) Representative images of Masson’s trichrome, picrosirius red, and van Gieson staining of wounds (n = 5 mice/group) 12 d after wounding are shown (n = 3 independent experiments). Bars: (A, C, and F) 100 µm; (B) 500 µm. (G) Hydroxyproline levels were measured in wounds at 12 d after wounding (**, P < 0.005; ***, P < 0.0005; n = 5 mice/group). (H) Quantitative TissueFAXS analyses of immunohistochemical staining shown in C are presented for keratinocytes (left) and fibroblasts (right; *, P < 0.05; **, P < 0.005; ***, P < 0.0005; n = 3 independent experiments). Means ± SD.