Hidehiko Baba, Hideyo Uchiwa, Shinichi Watanabe

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UVB Irradiation Increases the Release of SCF from Human Epidermal Cells Hidehiko Baba, Hideyo Uchiwa, Shinichi Watanabe Journal of Investigative Dermatology Volume 124, Issue 5, Pages 1075-1077 (May 2005) DOI: 10.1111/j.0022-202X.2004.23447.x Copyright © 2005 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 1 UVB stimulation increases SCF release from cultured human keratinocytes. (a) Effects of UVB irradiation on SCF release and cell viability of keratinocytes. The bars and polygonal line show the SCF content in culture medium and cell viability, respectively. SCF content was determined using an ELISA (Quantikine, R&D Systems, Minneapolis, Minnesota). Cell viability was measured using an alamarBlue Assay (Alamar Biosciences, Sacramento, California) 24 h after the second UVB irradiation. Each value is presented as the mean±SD of three determinations. **p<0.01, *p<0.05 as compared with the control by Dunnett's test. (b) Detection of soluble SCF by western blotting. Samples were concentrated 10-fold using a Centricon YM-10 (Millipore, Bedford, Massachusetts) and were separated by gel electrophoresis. Rabbit anti-SCF (Immuno-Biological Laboratories, Gunma, Japan) was used as a primary antibody. Journal of Investigative Dermatology 2005 124, 1075-1077DOI: (10.1111/j.0022-202X.2004.23447.x) Copyright © 2005 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 2 Changes of immunoreactivity toward SCF/c-kit in human epidermis following UVB exposure. Immunostaining for SCF (a–c) and c-kit (d–f) at pre-exposure (a, d), and 3 (b, e) and 10 days (c, f) after exposure. Acetone fixed-cryostat 5-μm sections were incubated with polyclonal rabbit anti-SCF (Santa Cruz Biotechnology, California) or a polyclonal rabbit anti-c-kit (DAKO, Carpinteria, California). Immunolocalization of the primary antibody was performed using the avidin–biotin peroxidase complex method (ABC-Elite kit, Vector Laboratories, Burlingame, California), with nickel-diaminobenzidine as the substrate. Journal of Investigative Dermatology 2005 124, 1075-1077DOI: (10.1111/j.0022-202X.2004.23447.x) Copyright © 2005 The Society for Investigative Dermatology, Inc Terms and Conditions