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

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© 2015 Lee et al.

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Figure 5. — Bosentan alleviates the accelerated wound-healing phenotype of CXXC5^−/− mice. After generation of full-thickness wounds (diameter = 1.5 cm) on the backs of CXXC5^+/+ and CXXC5^−/− mice, the mice were orally administered 100 mg/kg/d bosentan monohydrate daily for 11 d (n = 10 mice/group). (A) Representative images of macroscopic wounds, H&E staining, and IHC staining for β-catenin, CXXC5, keratin 14, collagen I, and PCNA in the wounds of CXXC5^+/+ and CXXC5^−/− mice treated or untreated with bosentan at 12 d after wounding (n = 4 mice/group) are shown (n = 3 independent experiments). Dashed lines indicate the epidermal–dermal boundary. F, fibroblasts; K, keratinocytes. (B) Relative wound closure rates after bosentan treatment in CXXC5^+/+ and CXXC5^−/− mice are 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 = 10 mice/group). (C) Western blot analyses of wound tissue lysates from CXXC5^+/+ and CXXC5^−/− mice treated with or without bosentan 12 d after wounding (n = 2 mice/group) were performed to detect β-catenin, CXXC5, keratin 14, α-SMA, collagen I, PCNA, endothelin-1, and Erk (n = 2 independent experiments). Relative densitometric ratios of each protein to Erk protein are shown. (D) Representative images of Masson’s trichrome, picrosirius red, or van Gieson staining of wound tissues obtained from CXXC5^+/+ and CXXC5^−/− mice treated with or without bosentan 12 d after wounding (n = 4 mice/group) are shown (n = 3 independent experiments). (A and D) Bars, 100 µm. (E) Hydroxyproline levels in wounds of CXXC5^+/+ and CXXC5^−/− mice treated with or without bosentan 12 d after wounding are shown (*, P < 0.05; ***, P < 0.0005; n = 4 mice/group). (F) Quantitative TissueFAXS analyses of IHC staining shown in A were performed in keratinocytes (left) and fibroblasts (right; *, P < 0.05; **, P < 0.005; ***, P < 0.0005; n = 3 independent experiments). Means ± SD.