1. TSLP Is a Potential Initiator of Collagen Synthesis and an Activator of CXCR4/SDF-1 Axis in Keloid Pathogenesis 
Jung U Shin, Seo Hyeong Kim, Hyeran Kim, Ji Yeon Noh, Shan Jin, Chang Ook Park, Won Jai Lee, Dong Won Lee, Ju Hee Lee, Kwang Hoon Lee 
Journal of Investigative Dermatology 
Volume 136, Issue 2, Pages (February 2016)
DOI: /j.jid
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2. Figure 1
Increased thymic stromal lymphopoietin (TSLP) expression in keloids and up-regulated collagen I, collagen III, and transforming growth factor-β (TGF-β) in fibroblasts after TSLP treatment. (a) Immunohistochemical staining of TSLP in keloids and normal skin. Bar = 200 μm. (b) Increased expression of TSLP was quantitatively measured using MetaMorph image analysis software (Universal Image Corp.). Data are representative of three normal tissues and three keloid tissues. Three images were taken from each tissue, and the mean staining intensity of each tissue was compared between normal and keloid tissues. Data are displayed as mean ± standard deviation SD. **P < (c) Real-time PCR analyses of collagen I, collagen III, and TGF-β in normal and keloid fibroblasts after treatment for 24 hours with 50 ng/ml TSLP. Results are displayed as mean ± standard deviation of two sets of experiments in triplicates. (d) Western blot analyses of collagen I, collagen III, and TGF-β in normal and keloid fibroblasts after treatment for 24 hours with 50 ng/ml TSLP. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) level was used as a loading control for western blots. (e) Release of collagen I, collagen III, and TGF-β in supernatants after treatment with 50 ng/ml TSLP measured using western blot analyses. (f) Western blot analyses of TSLP in normal fibroblasts and normal keratinocytes after treatment for 48 hours with 10 ng/ml TGF-β1. GAPDH level was used as a loading control for western blots. KF, keloid fibroblast; NF, normal fibroblast.
Journal of Investigative Dermatology  , DOI: ( /j.jid )
Copyright © 2015 The Authors Terms and Conditions

3. Figure 2
Up-regulated stromal cell-derived factor (SDF)-1α in normal fibroblasts and keloid fibroblasts after thymic stromal lymphopoietin (TSLP) treatment. (a) Western blot analyses of SDF-1α in normal fibroblasts and keloid fibroblasts after treatment for 24 hours with 50 ng/ml TSLP. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) level was used as a loading control for western blots. (b) SDF-1α release in the supernatant was measured after treatment with 50 ng/ml TSLP. (c) Immunohistochemical staining of SDF-1α in keloids and normal skin. Bar = 200 μm. (d) Increased expression of SDF-1α was quantitatively measured using MetaMorph image analysis software (Universal Image Corp.), and this increase was significant in keloids. Data are representative of three normal tissues and three keloid tissues. Three images were taken from each tissue, and the mean staining intensity of each tissue was compared between normal and keloid tissues. **P < KF, keloid fibroblast; NF, normal fibroblast.
Journal of Investigative Dermatology  , DOI: ( /j.jid )
Copyright © 2015 The Authors Terms and Conditions

4. Figure 3
Stromal cell-derived factor (SDF)-1α is expressed and colocalized in α-smooth muscle actin–positive (α-SMA+) myofibroblasts in keloid tissue. Three-color immunofluorescence confocal images were obtained for SDF-1α (green), α-SMA (red), and nuclei (blue) in (a) normal skin and (b) keloids. The images were merged, and double-positive cells are shown in yellow. Data are representative of experiments with three normal tissues and three keloid tissues. Original magnification ×400. Bar = 50 μm.
Journal of Investigative Dermatology  , DOI: ( /j.jid )
Copyright © 2015 The Authors Terms and Conditions

5. Figure 4
Increased CXCR4+ fibrocyte infiltration in keloid tissue. Three-color immunofluorescence confocal images were obtained for CXCR4 (green), procollagen-I (red), and CD45 (blue) in (a) normal skin and (b) keloids. Original magnification ×200. Bar = 50 μm. The image is representative of at least three skin lesions. (c) The images were merged, and double-positive cells are shown in yellow (CXCR4 + procollagen-I), magenta (procollagen-I + CD45), and cyan (CD45 + CXCR4). Original magnification ×400. Bar = 25 μm. (d) Middle and right panels represent keloid tissue. The overlay of CXCR4, procollagen-I, and CD45 is observed by increasing magnification: 400× and 630× (middle panel bar = 25 μm; right panel bar = 20 μm, respectively). The left panel represents normal skin staining. Original magnification ×200. Bar = 50 μm. The image is representative data of three normal tissues and three keloid tissues.
Journal of Investigative Dermatology  , DOI: ( /j.jid )
Copyright © 2015 The Authors Terms and Conditions

6. Figure 5
Increased stromal cell-derived factor-1α expression and CXCR4+ fibrocyte infiltration after intradermal thymic stromal lymphopoietin (TSLP) injection. (a) Mice (n = 4/group) received TSLP (10 μg) or mouse serum albumin (MSA; 10 μg) injections three times per week for 2 weeks. (b) Dermal thicknesses of MSA-treated and TSLP-treated mice were measured in hematoxylin and eosin staining. Results are displayed as mean ± standard deviation for each of the two groups. Original magnification ×200. Bar = 200 μm. *P < (c) Immunohistochemical staining of stromal cell-derived factor-1α in TSLP- or MSA-injected skin. Original magnification ×200 (left panel) and ×400 (right panel). Bar = 200 μm. (d–f) Three-color immunofluorescence confocal images were obtained for CXCR4 (green), procollagen-I (red), and CD45 (blue) in (d) MSA-injected and (e) TSLP-injected skin. Images are representative of experiments performed on eight mice. Bar = 50 μm. (f) The images were merged. The middle and right panels represent TSLP-injected skin. The overlay of CXCR4, procollagen-I, and CD45 is observed by increasing magnification: ×200 and ×400 (middle panel bar = 50 μm; right panel bar = 25 μm, respectively). The left panel represents MSA-injected skin staining, Original magnification ×200. Bar = 50 μm.
Journal of Investigative Dermatology  , DOI: ( /j.jid )
Copyright © 2015 The Authors Terms and Conditions