1. Effects of the Aminophenol Analogue p-Dodecylaminophenol on Mouse Skin
Noriko Takahashi, Yasunori Fujiu 
Journal of Investigative Dermatology 
Volume 130, Issue 5, Pages (May 2010)
DOI: /jid
Copyright © 2010 The Society for Investigative Dermatology, Inc Terms and Conditions

2. Figure 1
Skin irritation in RA-treated and p-DDAP-treated mouse skin. Mouse skin was treated without (control) or with RA (a) or p-DDAP (b) daily for 4 days. Changes in mouse skin were observed macroscopically.
Journal of Investigative Dermatology  , DOI: ( /jid )
Copyright © 2010 The Society for Investigative Dermatology, Inc Terms and Conditions

3. Figure 2
Expression of cytokines in mouse skin treated with RA and p-DDAP. Mouse skin was treated without (control, a) or with RA (b) or p-DDAP (c) daily for 4 days. Total RNA (5μg) was prepared, and reverse transcription PCR was performed using specific primers against each cytokine. After agarose gel (3%) electrophoresis, PCR products were analyzed by scanning densitometry. The expression levels of cytokine mRNA were normalized to that of glyceraldehyde-3-phosphate dehydrogenase. The cytokine expression of control was defined as 1.0. Results represent the mean±SD of each group (n=4). *P<0.05 versus control compared by Student's t-test.
Journal of Investigative Dermatology  , DOI: ( /jid )
Copyright © 2010 The Society for Investigative Dermatology, Inc Terms and Conditions

4. Figure 3
Histological features of RA-treated and p-DDAP-treated mouse skin. Mouse skin was treated without (control) or with RA (a) or p-DDAP (b) daily for 2 weeks. At the end of the treatment period, tissues were fixed in 4% buffered formalin and examined using light microscopy after sectioning and staining with hematoxylin and eosin. Skin epidermal thickness was assessed quantitatively. Each bar represents the mean±SD (n=6). *P<0.05 versus control compared by Student’s t-test.
Journal of Investigative Dermatology  , DOI: ( /jid )
Copyright © 2010 The Society for Investigative Dermatology, Inc Terms and Conditions

5. Figure 4
Gelatin zymography of MMPs in RA- and p-DDAP-treated mouse skin. Protein extracts (4μg) prepared from mouse skin treated without (control) or with RA or p-DDAP were applied to gelatin-impregnated gels. After electrophoresis, gels were incubated for 16hours at 37°C in 50mM Tris-HCl (pH 7.6) containing 5mM CaCl2, 0.2M NaCl, and 1μM ZnCl2. (a) Typical gelatin zymographic patterns were observed. (b) MMPs bands were densitometrically quantified by computer image analysis. The intensities of control bands were defined as 100%. Each bar represents the mean±SD (n=6). *P<0.05 versus control compared by Student's t-test.
Journal of Investigative Dermatology  , DOI: ( /jid )
Copyright © 2010 The Society for Investigative Dermatology, Inc Terms and Conditions

6. Figure 5
Comparison of HAase activities in mouse skin treated with RA or p-DDAP. HAase activities were measured in mouse skin treated without (control) or with RA (a) or p-DDAP (b). One unit of HAase activity was defined as the amount of enzyme required to produce 1μmol of reducing terminal GlcNAc per minute under the specified conditions. Each bar represents the mean±SD (n=6). *P<0.05 versus control compared by Student’s t-test.
Journal of Investigative Dermatology  , DOI: ( /jid )
Copyright © 2010 The Society for Investigative Dermatology, Inc Terms and Conditions

7. Figure 6
Comparison of 2D-PAGE patterns of proteins in RA-treated and p-DDAP-treated mouse skin. Proteins (150μg) prepared from mouse skin treated without (control) or with RA (a) or p-DDAP (b) were separated by 2D-PAGE with Immobiline DryStrip (pH 4–7, 18cm) and 10% 1D-PAGE and stained using SYPRO Ruby. Gel patterns shown are representative of five gels in each condition. Intensities of protein spots were analyzed by computer image analysis. Each bar represents the mean±SD. *P<0.05 versus control compared by Student’s t-test.
Journal of Investigative Dermatology  , DOI: ( /jid )
Copyright © 2010 The Society for Investigative Dermatology, Inc Terms and Conditions

8. Figure 7
Mass spectra of upregulated or unchanged protein in RA-treated mouse skin. Upregulated protein (Figure 6, proteins 1 and 6) and unchanged protein (Figure 6, protein a) were degraded by trypsin and analyzed by MALDI-TOF MS. Proteins 1 and 6 were identified as cytokeratin 16 (a), and protein a was identified as cytokeratin 10 (b). The data are disclosed elsewhere.
Journal of Investigative Dermatology  , DOI: ( /jid )
Copyright © 2010 The Society for Investigative Dermatology, Inc Terms and Conditions

9. Figure 8
Transcriptional activity of RA, 9-cis RA, and p-DDAP. Cells were transfected with expression plasmids for RARα, RARβ, RARγ, RXRα, RXRβ, or RXRγ, the luciferase reporter plasmid, and the control Renilla luciferase expression plasmid. Cells were treated with RA, 9-cis RA, and p-DDAP, and luciferase activities were measured. The activities of 1μM RA or 1μM 9-cis RA were defined as 100%. Each bar represents the mean±SE (n=3). a: DMSO, b: 0.1μM RA, c: 1μM RA, d: 0.5μM p-DDAP, e: 1 μM p-DDAP, f: 2μM p-DDAP, g: 1μM 9-cis RA.
Journal of Investigative Dermatology  , DOI: ( /jid )
Copyright © 2010 The Society for Investigative Dermatology, Inc Terms and Conditions