Localized Inflammatory Skin Disease Following Inducible Ablation of I Kappa B Kinase 2 in Murine Epidermis Athanasios Stratis, Manolis Pasparakis, Doreen.

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Localized Inflammatory Skin Disease Following Inducible Ablation of I Kappa B Kinase 2 in Murine Epidermis Athanasios Stratis, Manolis Pasparakis, Doreen Markur, Renate Knaup, Ruth Pofahl, Daniel Metzger, Pierre Chambon, Thomas Krieg, Ingo Haase Journal of Investigative Dermatology Volume 126, Issue 3, Pages 614-620 (March 2006) DOI: 10.1038/sj.jid.5700092 Copyright © 2006 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 1 Macroscopic phenotype of mice with spontaneous and Tamoxifen-induced deletion of IKK2 in the epidermis. Phenotype of K14-Cre-ERT2IKK2fl/fl mice (a) with spontaneous epidermis-specific deletion of IKK2 at 6 months and (b) 5 weeks after Tamoxifen-induced deletion. (c–e) DNA isolated from (c) various body sites or (d) from epidermis of the ear that was separated from the dermis of mice with spontaneous deletion at 6 months (KO) or controls, and (e) from a Tamoxifen-treated area 3 weeks after treatment was subjected to PCR analysis. Deletion is detected by the presence of a PCR band of 652bp; D. The lower band (533bp; FL) indicates the floxed allele. (c, e) DNA isolated from tail snips of the corresponding mice 3 weeks after birth served as controls. (d) The faint band that can be detected at the expected size of the fragment from the floxed allele is likely to result from invading immune cells. Journal of Investigative Dermatology 2006 126, 614-620DOI: (10.1038/sj.jid.5700092) Copyright © 2006 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 2 Inflammatory skin disease in mice with localized epidermis-specific deletion of IKK2. Histopathological phenotype of K14-Cre-ERT2IKK2fl/fl mice. Skin sections from Tamoxifen-induced mice (a) 3 weeks; or (c, e, i, k) 5 weeks after induction, (b, d, f, g, h) mice with spontaneous deletion of IKK2, and (l) control mice. Skin sections (a)–(h) were stained with hematoxylin/eosin. Sections (i)–(l) were immunostained with (i) antibodies recognizing macrophages (F4/80), (j) granulocytes (Gr-1), and (k, l) T cells (CD3). These sections were counterstained with propidium iodide to visualize tissue structure and nuclei. Bars: a, 100μm; b–e, 200μm; f, 20μm; g, h, 50μm; and i–l, 40μm. Journal of Investigative Dermatology 2006 126, 614-620DOI: (10.1038/sj.jid.5700092) Copyright © 2006 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 3 Analysis of epidermal differentiation markers in mice with epidermis-specific deletion of IKK2. Skin sections from K14-Cre-ERT2IKK2fl/fl mice spontaneously induced (left panel), tamoxifen-induced (middle panel), and from control mice (right panel) were immunostained for the indicated epidermal differentiation markers (green staining): keratin 14 (K14), keratin 10 (K10), filaggrin (Fil), and loricrin (Lor). Sections were counterstained with propidium iodide to visualize tissue structure and nuclei. Bar=40μm. Journal of Investigative Dermatology 2006 126, 614-620DOI: (10.1038/sj.jid.5700092) Copyright © 2006 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 4 Hair follicle disruption and signal transduction in the skin of K14-Cre-ERT2IKK2fl/fl mice. Confocal images of skin sections from (c, e, g, i, j) tamoxifen-induced and (a, b, d, m) spontaneously induced K14-Cre-ERT2IKK2fl/fl mice and (f, h, k, l, n) controls. The green signal shows immunostaining against (a) keratin 14, (b) keratin 10, (c) filaggrin, (d–f) keratin 6, (g, h) CCAAT displacement enhancer protein, (i, l) laminin 5, (j, k) phosphorylated signal transducer and activator of transcription 3 (STAT3), and (m, n) phosphorylated extracellular signal-regulated kinase 1/2 (ERK1/2). Sections (a)–(i) and (l) were counterstained with propidium iodide to visualize tissue structure and nuclei. Inserts in (k) and (n) show high-power magnification of the indicated parts of the epidermis. Bars=40μm. Western blot analysis of ERK phosphorylation (o) of affected skin samples from K14-Cre-ERT2IKK2fl/fl mice (KO) and of wild-type skin samples (WT). The upper panel shows phosphorylated ERK (pERK), and the lower panel shows ERK1/2 as loading control (ERK1/2). Journal of Investigative Dermatology 2006 126, 614-620DOI: (10.1038/sj.jid.5700092) Copyright © 2006 The Society for Investigative Dermatology, Inc Terms and Conditions