1. IL-31 Receptor Alpha Expression in Epidermal Keratinocytes Is Modulated by Cell Differentiation and Interferon Gamma 
Ruth Heise, Mark M. Neis, Yvonne Marquardt, Sylvia Joussen, Peter C. Heinrich, Hans F. Merk, Heike M. Hermanns, Jens M. Baron 
Journal of Investigative Dermatology 
Volume 129, Issue 1, Pages (January 2009)
DOI: /jid
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2. Figure 1
mRNA expression of IL-31 Rα decreases in differentiated normal human epidermal keratinocytes (NHEK) and is upregulated after stimulation with IFNγ. TaqMan real-time PCR analysis of NHEK cultured in differentiation medium containing 1.4mM Ca2+: upregulation of (a) cytokeratin 10 (KRT 10) and (b) loricrin (LOR) mRNA expression serves as marker for the induced differentiation process. (c) Expression of IL-31Rα mRNA was 29-fold reduced in differentiated keratinocytes (day 15) compared with proliferating keratinocytes (d 0). (d) Expression of oncostatin M receptor beta mRNA was decreased on days 1 and 5 but not significantly altered during the late phases of keratinocyte differentiation. The relative mRNA levels were normalized to 18S rRNA. (e) Expression of IL-31Rα mRNA was fivefold induced in proliferating NHEK 24h after stimulation with 20ngml-1 INF-γ. Incubation of NHEK with 10ngml-1 IL-1α, 10ngml-1 TNF-α and a cocktail containing 10ngml-1 IL-1α, 5ngml-1 TNF-α, and 5ngml-1 IL-6 for 24h did not alter IL-31Rα mRNA expression. (f) Twenty four hours after stimulation with 20ngml-1 INF-γ, expression of IL-31Rα mRNA was 3.8-fold induced in differentiated NHEK (cultured in medium containing 1.4mM Ca2+ for 8 days). Incubation of NHEK with 10ngml-1 IL-1α, 10ngml-1 TNF-α, and a cocktail containing 10ngml-1 IL-1α, 5ngml-1 TNF-α, and 5ngml-1 IL-6 for 24h did not alter IL-31Rα mRNA expression.
Journal of Investigative Dermatology  , DOI: ( /jid )
Copyright © 2009 The Society for Investigative Dermatology, Inc Terms and Conditions

3. Figure 2
Influence of cell differentiation and inflammatory cytokines on IL-31 Rα protein expression and IL-31 Rα dependent signaling pathways. Western blot analysis of IL-31Rα expression in NHEK (a) at different stages of cell differentiation, (b) after stimulation with pro-inflammatory cytokines. (c) Analysis of signaling pathways in nondifferentiated (d 0) and differentiated (d 15) NHEK in response to oncostatin M and IL-31. (a) For Western blot analysis, NHEK were cultured in differentiation medium containing 1.4mM Ca2+. Cells were harvested at the times indicated and total cell lysates were prepared and separated by SDS-PAGE. = histological features of NHEK at day 0 or 15. Dermal fibroblasts (NHDF) and monocytes (MØ) stimulated with 1000Uml-1 IFN-γ for 48h were used as negative or positive controls. (b) Proliferating NHEK were incubated with 10ngml-1 IL-1α, 10ngml-1 TNF-α, 20ngml-1 INF-γ, and a cocktail containing 10ngml-1 IL-1α, 5ngml-1 TNF-α and 5ngml-1 IL-6 for 48h. Western blots were developed with an antibody against IL-31Rα. Blots were stripped and reprobed with a primary monoclonal anti-human β-actin antibody. (c) NHEK at day 0 or cultured in differentiation medium containing 1.4mM Ca2+ for 15 days were stimulated with 20ngml-1 OSM or 100ngml-1 IL-31 for the indicated time points. Total cell lysates were separated by SDS-PAGE. Western blots were developed with antisera specific for the indicated proteins.
Journal of Investigative Dermatology  , DOI: ( /jid )
Copyright © 2009 The Society for Investigative Dermatology, Inc Terms and Conditions