The Epidermal Stem Cell Factor Is Over-Expressed in Lentigo Senilis: Implication for the Mechanism of Hyperpigmentation Hideko Hattori, Makoto Kawashima,

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The Epidermal Stem Cell Factor Is Over-Expressed in Lentigo Senilis: Implication for the Mechanism of Hyperpigmentation Hideko Hattori, Makoto Kawashima, Yoshiaki Ichikawa, Genji Imokawa, PhD Journal of Investigative Dermatology Volume 122, Issue 5, Pages 1256-1265 (May 2004) DOI: 10.1111/j.0022-202X.2004.22503.x Copyright © 2004 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 1 Clinical appearance (A) and histology (B, C, and D) of lentigo senilis (LS). (A) Clinical appearance of LS (Patient 1) in the upper arm of a Japanese female aged 70 y; there is an increased basal pigmentation and a slight acanthosis with prolonged rete ridges in the lesional epidermis in LS. (B) Hematoxylin and eosin (H&E) staining, × 50 (Patient 1). (C) Toluidine blue staining in the lesion, × 50 (Patient 27). (D) Toluidine blue staining in the non-lesion, × 50 (Patient 27). Journal of Investigative Dermatology 2004 122, 1256-1265DOI: (10.1111/j.0022-202X.2004.22503.x) Copyright © 2004 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 2 RT-PCR analysis of stem cell factor (SCF) mRNA in the epidermis of lentigo senilis (LS) lesions and in perilesional skin. (A) Fluorogram, (B) densitometric analysis. NL: non-lesional epidermis; L: lesional epidermis of LS. Fluorograms are shown at 34 or 40 cycles of PCR for SCF and at 22 or 35 cycles for glyceraldehyde-3-phosphate dehydrogenase (G3PDH). **p<0.01, *p<0.05. Journal of Investigative Dermatology 2004 122, 1256-1265DOI: (10.1111/j.0022-202X.2004.22503.x) Copyright © 2004 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 3 RT-PCR analysis of c-kit mRNA in the epidermis of lentigo senilis (LS) lesions and in perilesional skin. (A) Fluorogram, (B) densitometric analysis. NL: Non-lesional epidermis; L: lesional epidermis of LS. Fluorograms are shown at 35 or 40 cycles of PCR for c-KIT and at 25 or 35 cycles for glyceraldehyde-3-phosphate dehydrogenase (G3PDH). NS: not significant. Journal of Investigative Dermatology 2004 122, 1256-1265DOI: (10.1111/j.0022-202X.2004.22503.x) Copyright © 2004 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 4 RT-PCR analysis of growth-related oncogene α (GROα) mRNA in lentigo senilis (LS) lesions and in perilesional skin. (A) Fluorogram, (B) densitometric analysis. NL: Non-lesional epidermis; L: lesional epidermis of LS. Fluorograms are shown at 31 cycles of PCR for GROα and at 22 cycles for glyceraldehyde-3-phosphate dehydrogenase (G3PDH). NS: not significant. Journal of Investigative Dermatology 2004 122, 1256-1265DOI: (10.1111/j.0022-202X.2004.22503.x) Copyright © 2004 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 5 RT-PCR analysis of various transcripts used in the epidermis of the same donor with lentigo senilis (LS) and perilesional skin. NL: Non-lesional epidermis; L: lesional epidermis of LS. Fluorograms are shown at 45, 40, 40, 40, 40, and 35 cycles of PCR for ET-1, endothelin B receptor (ETBR), tyrosinase, stem cell factor (SCF), c-KIT, and glyceraldehyde-3-phosphate dehydrogenase (G3PDH), respectively. The numbers above the bands represent the densitometric intensities of those bands. Journal of Investigative Dermatology 2004 122, 1256-1265DOI: (10.1111/j.0022-202X.2004.22503.x) Copyright © 2004 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 6 Immunohistochemistry with an antibody to stem cell factor (SCF) in the epidermis of a lentigo senilis (LS) lesion: Patient 20 (A) Lesional skin (× 100), (B) non-lesional skin (× 100). Journal of Investigative Dermatology 2004 122, 1256-1265DOI: (10.1111/j.0022-202X.2004.22503.x) Copyright © 2004 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 7 Immunostaining with an endothelin-1 (ET-1) antibody in the epidermis of the same donor with lentigo senilis (LS) as used for stem cell factor (SCF) immunostaining (inFigure 6). Patient 20: (A) Lesional skin; (B) non-lesional skin (× 100). Journal of Investigative Dermatology 2004 122, 1256-1265DOI: (10.1111/j.0022-202X.2004.22503.x) Copyright © 2004 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 8 Western blotting of stem cell factor (SCF) protein in the epidermis of lentigo senilis (LS) lesions. (A) A representative fluorogram in western blotting, (B) fluorogram of six LS skins, (C) densitometric analysis of (B). Extracts of separated epidermis from LS lesions (5 μg protein per lane) were subjected to 16.5% (A) or 15% (B) acrylamide gel electrophoresis, followed by electroblotting and then immunostaining with antibodies to SCF as noted. The protein staining was concomitantly performed with Coomassie Brilliant Blue according to standard procedures. **p<0.01, *p<0.05; NHKC: UVB (40 mJ per cm2) exposed normal human keratinocytes. Cell lysate was taken 24 h after UVB radiation; recSCF: recombinant SCF. Journal of Investigative Dermatology 2004 122, 1256-1265DOI: (10.1111/j.0022-202X.2004.22503.x) Copyright © 2004 The Society for Investigative Dermatology, Inc Terms and Conditions