1. Cartilage Oligomeric Matrix Protein Increases in Photodamaged Skin
Masaki Kobayashi, Keigo Kawabata, Ayumi Kusaka-Kikushima, Yoshinori Sugiyama, Tomotaka Mabuchi, Susumu Takekoshi, Muneo Miyasaka, Akira Ozawa, Shingo Sakai 
Journal of Investigative Dermatology 
Volume 136, Issue 6, Pages (June 2016)
DOI: /j.jid
Copyright © 2016 The Authors Terms and Conditions

2. Figure 1
COMP mRNA expression is higher in preauricular skin than in postauricular skin. (a) COMP mRNA was measured by quantitative real-time PCR in total RNA extracted from postauricular and preauricular skin of the same subjects (n = 8). Glyceraldehyde 3-phophate dehydrogenase was used as an internal control to normalize gene expression. *P < (b) COMP-mRNA-positive cells were detected in pre/postauricular skin by in situ hybridization using antisense probes (upper) and sense probes (lower). Bar = 50 μm. COMP, cartilage oligomeric matrix protein.
Journal of Investigative Dermatology  , DOI: ( /j.jid )
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3. Figure 2
The localization and expression of COMP in sun-exposed skins vary with the progress of photoaging. (a) The expression of COMP mRNA in fibroblasts was detected by in situ hybridization (arrow). (b) The number of COMP-mRNA-positive cells was counted at each stage using index by Kawabata et al. (2014). Graph data represent the mean ± SEM per 6 × field of 3–5 subjects. *P < 0.05 (versus stage III). (c) The expression of COMP protein was detected by immunohistochemical staining. The photographs are representative of each stage. (d) The intensity of COMP signals was evaluated by five blind scorers using a 10-cm visual analog scale. Graph data represent the mean ± SD. *P < 0.05, ***P < COMP, cartilage oligomeric matrix protein; SD, standard deviation; SEM, standard error of the mean.
Journal of Investigative Dermatology  , DOI: ( /j.jid )
Copyright © 2016 The Authors Terms and Conditions

4. Figure 3
COMP colocalizes with collagen fibrils and amorphous elastotic materials in photoaged dermis. The papillary dermis, the area of the dermal-epidermal junction (a), the reticular dermis in the photodamaged skin (stage V (c)), and the photo-protected skin (e) were observed in electron micrographs. Immunolocalization of COMP was visualized with immunoperoxidase-diaminobenzidine before embedding. Electron micrographs at high magnification showed (b) collagen bundles beneath the epidermis and (d) collagen bundles and amorphous materials in the area of solar elastosis, where COMP signals were detected, and showed (f) normal elastic fiber in the reticular dermis of sun-protected skin. Arrowhead, COMP signals; asterisk, collagen fibrils; epi, epidermis; sharp, elastotic materials. Bar = 5 μm. Representative data from four subjects are shown. COMP, cartilage oligomeric matrix protein.
Journal of Investigative Dermatology  , DOI: ( /j.jid )
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5. Figure 4
TGF-β takes part in the UVA-induced increase of COMP. (a) COMP protein was analyzed by western blotting with cultured media of UVA-irradiated fibroblasts. (b) COMP mRNA was analyzed by real-time PCR 24 hours after UVA irradiation. Glyceraldehyde 3-phophate dehydrogenase was used as an internal control to normalize gene expression. Each value represents the mean ± SD. *P < 0.05, **P < 0.01 versus 0 J/cm2. The effect of an anti-TGF-β neutralizing antibody (1 μg/ml) on protein and mRNA expression was assessed by western blotting (c) and real-time PCR (d). The effect of SB on protein and mRNA expression was assessed by western blotting (e) and real-time PCR (f). Graph data represent the mean ± SD. **P < 0.01. Loading was controlled by staining membrane with Amido Black (Supplementary Figure S3 online). COMP, cartilage oligomeric matrix protein; SD, standard deviation; TGF-β, transforming growth factor- β; UVA, ultraviolet A.
Journal of Investigative Dermatology  , DOI: ( /j.jid )
Copyright © 2016 The Authors Terms and Conditions

6. Figure 5
UVA irradiation elicits various effects on TGF-β isoforms expression in cultured NHDFs. (a) TGF-β1, (b) TGF-β2, and (c) TGF-β3 were analyzed by real-time PCR after UVA irradiation. Each value represents the mean of a ratio (expression in UVA-treated cells/expression in UVA-untreated cells) ± SD, *P < 0.05, ***P < versus 0 J/cm2. (d) Total and active TGF-β1 protein was analyzed by the ELISA kit. Graph data represent the mean ± SD, *P < 0.05, #P = 0.06 versus control. SD, standard deviation; TGF-β, transforming growth factor- β; UVA, ultraviolet A.
Journal of Investigative Dermatology  , DOI: ( /j.jid )
Copyright © 2016 The Authors Terms and Conditions