1. The Effects of Dickkopf 1 on Gene Expression and Wnt Signaling by Melanocytes: Mechanisms Underlying Its Suppression of Melanocyte Function and Proliferation 
Yuji Yamaguchi, Thierry Passeron, Hidenori Watabe, Ken-ichi Yasumoto, Francois Rouzaud, Toshihiko Hoashi, Vincent J. Hearing 
Journal of Investigative Dermatology 
Volume 127, Issue 5, Pages (May 2007)
DOI: /sj.jid
Copyright © 2007 The Society for Investigative Dermatology, Inc Terms and Conditions

2. Figure 1
DKK1 expression by palmoplantar and by non-palmoplantar fibroblasts. (a) Immunoblotting analysis; the intensity of the 35kDa DKK1 band is higher by 2.8-fold (P= ) in palmoplantar fibroblasts (Palm1, Palm2) than in non-palmoplantar fibroblasts (Trunk1, Trunk2). (b) Immunocytochemical analysis; high levels of DKK1 (green fluorescence) are seen in palmoplantar fibroblasts (Palm3) compared with non-palmoplantar fibroblasts (Trunk3). Nuclei are stained blue by 4′,6-diamidino-2-phenylindole. Bar=64μm.
Journal of Investigative Dermatology  , DOI: ( /sj.jid )
Copyright © 2007 The Society for Investigative Dermatology, Inc Terms and Conditions

3. Figure 2
Validation of gene expression levels using RT-PCR. Shown are normal human melanocytes treated with 50ng/ml DKK1 for 2hours or untreated (ctrl). The expression of PKCβ1 (P<0.0001), Krn1 (P<0.0001), GPR51 (P<0.01), LRP6 (P=0.065), LDLR (P<0.001), TNFR10 (P<0.0001), and Gadd45 (P<0.002) is upregulated in melanocytes when treated with DKK1, but MITF (P<0.001) is downregulated. glyceraldehyde-3-phosphate dehydrogenase expression served as control (P=0.41) (n>=3).
Journal of Investigative Dermatology  , DOI: ( /sj.jid )
Copyright © 2007 The Society for Investigative Dermatology, Inc Terms and Conditions

4. Figure 3
Validation of expression levels of proteins related to Wnt signaling pathways in normal human melanocytes treated with DKK1 for (a) 2hours or (b) 5 days or untreated (ctrl) by immunoblotting. GSK3β expression is unchanged at 2hours, but is downregulated at 5 days. The expression of Ser9-phosphorylated GSK3β and PKCα is upregulated, but β-catenin expression is downregulated. Numbers under the gels indicate levels of intensity compared with control.
Journal of Investigative Dermatology  , DOI: ( /sj.jid )
Copyright © 2007 The Society for Investigative Dermatology, Inc Terms and Conditions

5. Figure 4
Expression of pGSK3β, PKCα, and β-catenin by normal human melanocytes. Shown are normal human melanocytes treated with 50ng/ml DKK1 for 2hours or untreated (ctrl) by immunocytochemistry. The expression of Ser9-phosphorylated GSK3β and PKCα is upregulated in response to DKK1, but β-catenin expression is downregulated. Green fluorescence marks antibody staining, and nuclei are stained blue by 4′,6-diamidino-2-phenylindole. Bar=128μm for outer panels and 64μm for insets.
Journal of Investigative Dermatology  , DOI: ( /sj.jid )
Copyright © 2007 The Society for Investigative Dermatology, Inc Terms and Conditions

6. Figure 5
Immunostaining of GSKβ, Ser9-phosphorylated GSKβ, PKCα, and β-catenin in the reconstructed epidermis. Immunostaining shown after 10 days of treatment with or without 100ng/ml rhDKK1. Note the decreased expression of GSK3β (0.63) and β-catenin (DKK1/ctrl ratio=0.76) and the increased expression of Ser9-phosphorylated GSK3β (2.00) and PKCα (1.11).
Journal of Investigative Dermatology  , DOI: ( /sj.jid )
Copyright © 2007 The Society for Investigative Dermatology, Inc Terms and Conditions

7. Figure 6
Immunohistochemistry of GSKβ, PKCα, and β-catenin in palmoplantar and non-palmoplantar skin in situ. Melanocytes in the skin are identified by MART-1 staining (red fluorescence). Palm skin (left) shows low expression of GSK3β (Palm/Trunk ratio=0.59) but high expression of GSK3β phosphorylated at Ser9 (1.24) and of PKCα (2.73). In contrast, levels of β-catenin (0.02) expression are lower in palmoplantar melanocytes in situ compared with melanocytes in trunk skin (right). Bar=100μm for upper six panels and 24μm for lower two panels.
Journal of Investigative Dermatology  , DOI: ( /sj.jid )
Copyright © 2007 The Society for Investigative Dermatology, Inc Terms and Conditions