Keratinocyte Differentiation in Hyperproliferative Epidermis: Topical Application of PPARα Activators Restores Tissue Homeostasis László G. Kömüves,

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Keratinocyte Differentiation in Hyperproliferative Epidermis: Topical Application of PPARα Activators Restores Tissue Homeostasis László G. Kömüves, Karen Hanley, Mao-Qiang Man, Peter M. Elias, Mary L. Williams, Kenneth R. Feingold Journal of Investigative Dermatology Volume 115, Issue 3, Pages 361-367 (September 2000) DOI: 10.1046/j.1523-1747.2000.00076.x Copyright © 2000 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 1 Effects of topical clofibrate treatment on epidermal morphology. Epidermal hyperproliferation was produced by EFAD diet or by repeated barrier disruption in adult hairless mice. The animals were treated twice a day with vehicle or with clofibrate for 3 d. The treatment resulted in a significant decrease in epidermal thickness (see Table 1for quantitative data). Hematoxylin and eosin staining. Scale bar: 10 μm. Journal of Investigative Dermatology 2000 115, 361-367DOI: (10.1046/j.1523-1747.2000.00076.x) Copyright © 2000 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 2 Effects of topical PPAR& ALPHA; activator treatment on cell proliferation in hyperproliferative skin. Proliferative cells were detected in normal control hairless mice (A), or in hyperproliferative skin produced by EFAD diet (B, C, D), or by repeated barrier disruption (E, F, G) in adult hairless mice. The animals were treated twice a day with vehicle (B, E), clofibrate (C, F), or Wy-14,643 (D, G) for 3 d. Proliferating cells were identified by immunohistochemical detection of a cell-proliferation-specific marker, PCNA. The sections were counterstained with Sytox Green to detect nuclei (green color). Proliferating cells are identified by yellow nuclei, a result of overlaid green and red signals. Scale bars: 50 μm. Journal of Investigative Dermatology 2000 115, 361-367DOI: (10.1046/j.1523-1747.2000.00076.x) Copyright © 2000 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 3 Effects of topical clofibrate treatment on the localization of involucrin, profilaggrin-filaggrin, and loricrin proteins in hyperproliferative skin. Epidermal hyperproliferation was produced by EFAD diet (A-F) or by repeated barrier disruption (G-L) in adult hairless mice. The animals were then treated twice a day with vehicle (A-C, G-I) or with clofibrate (D-F, J-L) for 3 d. Mouse involucrin (A, D, G, J), profilaggrin-filaggrin (B, E, H, K), and loricrin (C, F, I, L) were detected using specific antibodies. For details of the immunolocalization see Materials and Methods. Scale bar: 20 μm. Journal of Investigative Dermatology 2000 115, 361-367DOI: (10.1046/j.1523-1747.2000.00076.x) Copyright © 2000 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 4 Effects of topical clofibrate treatment on the expression of profilaggrin and loricrin genes detected by in situ hybridization. Epidermal hyperproliferation was produced by EFAD diet (A-D) or by repeated barrier disruption (E-H). The animals were then treated twice a day with vehicle (A, B, E, F) or with clofibrate (C, D, G, H) for 3 d. Profilaggrin (A, C, E, G) and loricrin mRNA (B, D, F, H) was detected with digoxigenin-labeled riboprobes. Scale bar: 20 μm. Journal of Investigative Dermatology 2000 115, 361-367DOI: (10.1046/j.1523-1747.2000.00076.x) Copyright © 2000 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 5 Effects of topical PPAR&ALPHA; activator treatment on apoptosis. Apoptosis was detected in normal control hairless mice (A), or in hyperproliferative skin produced by EFAD diet (B, C, D), or by repeated barrier disruption (E, F, G) in adult hairless mice. The animals were treated twice a day with vehicle (B, E), clofibrate (C, F), or Wy-14,643 (D, G) for 3 d. Apoptotic cells were detected by the TUNEL assay, and the FITC-uridine label was visualized by confocal microscopy. The sections were counterstained with Sytox Orange to detect nuclei (red color). Apoptotic cells are identified by yellow-green nuclei. Scale bars: 50 μm. Journal of Investigative Dermatology 2000 115, 361-367DOI: (10.1046/j.1523-1747.2000.00076.x) Copyright © 2000 The Society for Investigative Dermatology, Inc Terms and Conditions