Fig 1.
E2F1-deficiency accelerates wound healing.
(A) Representative skin wounds in WT and E2F1–/– mice at serial time points post- surgery. (B) Quantifications of the wound healing rates (percent wound closure). *p<0.05 vs. WT at same time points. N.S., not significant; n = 10 per group.
Fig 2.
The dermal thickness and collagen amount are greater in E2F1–/– mice than in WT mice.
Wound tissues were isolated at day 7 post-surgery and analyzed histologically. (A-B) H.E. and (C) Masson’s trichrome staining. Both entire wound (A) and wound border zone (B-C) are shown. Arrowheads indicate edges of wound granulation tissue. Double-headed arrow bars indicate the distance between the leading edges of wounded epidermis. (D) Quantifications of Epithelial gap (left panel), dermal thickness (middle panel), and collagen deposition (right panel). **p<0.01, *p<0.05; n = 10 per group.
Fig 3.
E2F1-deficiency enhances vessel growth in the border area of the skin wound.
At day 7 post-surgery, Lectin was i.v. injected 10 min before euthanasia to identify vasculature; subsequent analyses were performed in sections of wound skin and quantified at the border zone of wounds. (A) Functional vessels were identified by staining with anti-lectin antibodies (red) (left and middle panels, scale bar = 50mm) and quantified (right panel). (B) Proliferation cells were identified by staining for BrdU (green) (left and middle panels) and quantified (right panel). (C) Proliferating ECs (yellow) were identified by co-staining for BrdU (green) and lectin (red) (left and middle panels, scale bar = 50 mm) and quantified as the number of lectin+BrdU+ vessels per mm2 (right panel); nuclei were stained with DAPI (blue). **p<0.01; n = 10 per group.
Fig 4.
More infiltrating macrophages are found in the wound of E2F1–/– mice than in WT mice.
At day 7 post-surgery, the wound skin tissues were analyzed immunochemically and by qRT-PCR. Shown are representative staining (left and middle panels) (original magnification, X400; 0.06 mm2/field) and quantifications (right panels) of the cells stained positive for CD68 (A), MPO (B) and CD3 (C). (D) The expression levels of M1 and M2 macrophage marker genes in the wounded skin tissues of WT and E2F1–/– mice were analyzed by qRT-PCR. **p<0.01. n.s., not significant; n = 10 per group.
Fig 5.
VEGF-A level is significantly higher in the wound of E2F1–/– mice than in WT mice.
Wound tissues were isolated at day 7 post-surgery. (A) Immumohistochemical staining of VEGF (brown), 20X original magnification (left panels), the inlets were enlarged (right panels). (B) VEGF mRNA expression was analyzed by qRT-PCR and normalized to the level of GAPDH. (C) Representatives of Western blotting (upper panel) and quantification of VEGF protein levels (lower panel). Protein levels were quantified densitometrically and normalized to tubulin levels. **p<0.01; n = 5.