1. Differential Contribution of Dermal Resident and Bone Marrow–Derived Cells to Collagen Production during Wound Healing and Fibrogenesis in Mice 
Reiichi Higashiyama, Sachie Nakao, Yayoi Shibusawa, Osamu Ishikawa, Tadashi Moro, Kenichiro Mikami, Hiroshi Fukumitsu, Yoshitaka Ueda, Kaori Minakawa, Yasuhiko Tabata, George Bou-Gharios, Yutaka Inagaki 
Journal of Investigative Dermatology 
Volume 131, Issue 2, Pages (January 2011)
DOI: /jid
Copyright © 2011 The Society for Investigative Dermatology, Inc Terms and Conditions

2. Figure 1
Identification of specific fluorescent signals in dermal tissue. The emission fingerprinting method was used to distinguish the specific enhanced green fluorescent protein (EGFP) signals from the background autofluorescence. Using a computer program installed in the confocal laser-scanning microscope, a mixture of specific and nonspecific fluorescent signals (a) can be separated into the specific EGFP fluorescence (green in b) and nonspecific autofluorescence (blue in b). Representative pictures are shown of dermal tissue obtained from four COL/EGFP recipient mice that were treated repeatedly with bleomycin (BLM) injections for 28 days. Arrows in b indicate the bone marrow (BM)-derived EGFP-positive cells that express collagen type I. Scale bars=50μm.
Journal of Investigative Dermatology  , DOI: ( /jid )
Copyright © 2011 The Society for Investigative Dermatology, Inc Terms and Conditions

3. Figure 2
Activation of Col1a2 promoter during dermal excision healing. Dermal specimens were obtained from transgenic COL/EGFP mice before an excision (a, e), or at day 2 (b, f, and i), day 7 (c, g, j), and day 14 (d, h) after wounding. They were subjected to Sirius red-Fast green FCF staining (a–d) or confocal microscopic examination (e–j) detecting enhanced green fluorescent protein (EGFP; green) and α-smooth muscle actin (αSMA; red) expression. Representative pictures are shown from four mice in each group. The wound margins and beds are indicated by arrowheads and hatched arrows, respectively, in the inflammatory (b, f, i) and proliferative (c, g, j) stages. Arrows indicate EGFP-positive αSMA-expressing cells. Scale bars=100μm (a–d) or 50μm (e–j).
Journal of Investigative Dermatology  , DOI: ( /jid )
Copyright © 2011 The Society for Investigative Dermatology, Inc Terms and Conditions

4. Figure 3
Activation of Col1a2 promoter in dermal resident cells during excision healing. Expression of enhanced green fluorescent protein (EGFP; green) was examined in dermal specimens obtained from CAG/EGFP recipient mice (a–d) or COL/EGFP recipients (e–h) before a dermal excision (a, e), or at day 7 (b, c, f, g) and day 14 (d, h), which correspond to the proliferative and remodeling stages of wound healing process, respectively. They were stained with specific antibodies recognizing α-smooth muscle actin (αSMA; red). Representative pictures are shown from four mice in each group. The wound margins and beds are indicated by arrowheads and hatched arrows, respectively, in the proliferative stage. Scale bars=50μm.
Journal of Investigative Dermatology  , DOI: ( /jid )
Copyright © 2011 The Society for Investigative Dermatology, Inc Terms and Conditions

5. Figure 4
Quantitative analysis of Col1a2 promoter activation during dermal excision healing. Transgenic COL/LUC mice (Tg) and their bone marrow (BM) recipients (Rc) were given dermal excisions. Skin specimens were obtained from those mice before the excision, or at days 7 and 14 after wounding. Luciferase assays were carried out to determine Col1a2 promoter activity. Samples obtained from wild-type (WT) mice were also analyzed as a control. Luciferase activity was normalized against the protein concentration of the tissue homogenates. The values are expressed as means±SD from 3 to 4 mice in each group. *Statistically significant difference between the groups (P=0.046). NS, not significant; RLU, relative luminescence units.
Journal of Investigative Dermatology  , DOI: ( /jid )
Copyright © 2011 The Society for Investigative Dermatology, Inc Terms and Conditions

6. Figure 5
Activation of Col1a2 promoter during dermal fibrogenesis. Dermal specimens of transgenic COL/EGFP mice were obtained 21 days after a single injection of control poly-L-lactic acid (PLA; a, c, e) or bleomycin (BLM)–PLA (b, d, f). They were subjected to hematoxylin and eosin (H&E; a, b) and Sirius red-Fast green FCF staining (c, d), or confocal microscopic examination (e, f) detecting enhanced green fluorescent protein (EGFP)-positive (green) and α-smooth muscle actin (αSMA)-positive (red) cells. Representative pictures are shown from four mice in each group. Arrows indicate EGFP-positive cells that coexpress αSMA. Under the panels are shown the mean numbers of EGFP-positive cells per area with the ratios of αSMA-expressing cells among them in parentheses. *Significantly higher value than that in the PLA-injected control (P=0.037). Scale bars=50μm.
Journal of Investigative Dermatology  , DOI: ( /jid )
Copyright © 2011 The Society for Investigative Dermatology, Inc Terms and Conditions

7. Figure 6
Activation of Col1a2 promoter in bone marrow (BM)-derived cells migrating into bleomycin (BLM)-induced fibrotic dermal tissue. Expression of enhanced green fluorescent protein (EGFP; green) was examined in dermal specimens obtained from CAG/EGFP recipient mice (a, c) or COL/EGFP recipients (b, d) 21 days after a single BLM–poly-L-lactic acid (PLA) injection. They were stained with specific antibodies recognizing either α-smooth muscle actin (αSMA; red in a, b) or CD45 (red in c, d). Representative pictures are shown from four mice in each group. Arrows in b and d indicate EGFP-positive BM-derived collagen-producing cells, whereas arrowheads in d show CD45-positive cells that do not produce collagen type I. Arrows in c represent CD45-positive BM-derived cells observed in CAG/EGFP recipient mice. Scale bars=50μm (a, b, and upper panels of c, d) or 10μm (lower panels of c, d).
Journal of Investigative Dermatology  , DOI: ( /jid )
Copyright © 2011 The Society for Investigative Dermatology, Inc Terms and Conditions