Regulation of IL-13 Receptors in Human Keratinocytes

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Regulation of IL-13 Receptors in Human Keratinocytes Rahul Purwar, Thomas Werfel, Miriam Wittmann Journal of Investigative Dermatology Volume 127, Issue 5, Pages 1271-1274 (May 2007) DOI: 10.1038/sj.jid.5700687 Copyright © 2007 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 1 Human KCs express functional IL-13Rα1. KCs were isolated as described previously (Purwar et al., 2006). (a) KCs were stimulated with indicated cytokine(s) for 24hours and IL-13Rα1 expression was determined by flow cytometry using an IL-13Rα1 mAb. Representative histogram is shown from six different experiments with cells from different donors. (b) KCs were stimulated with IL-13 (50ng/ml) or IL-4 (50ng/ml) for 15minutes. Cells were lysed and phospho-STAT6 was detected by Western blotting. (c and d) KCs were stimulated with TGF-β1 (1ng/ml) and IL-13 (50ng/ml) (24–72hours; Figure 1c: 72hours). CCL5 was measured in cell-free supernatants by CCL5-specific ELISA according to the manufacturer's instructions (Duoset, RnD systems). Mean±SEM is given. **P<0.02 and ***P<0.01 (compared to NS, not stimulated). Journal of Investigative Dermatology 2007 127, 1271-1274DOI: (10.1038/sj.jid.5700687) Copyright © 2007 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 2 Expression and cytokine regulation of IL-13Rα2 in KCs. KCs were stimulated with IL-13 (50ng/ml), IFN-γ (10ng/ml), TNF-α (10ng/ml) for 24hours (protein), or 6hours (RNA). (a) Representative dot-plots are shown from five different experiments with cells from different donors. The values shown in dot-plots correspond to percentage of IL-13Rα2-positive cells. (b) Total RNA was isolated and complementary DNA was synthesized. Data in bar graphs are shown as percentage induction of IL-13Rα2 messenger RNA after indicated cytokine stimulation. (c) Supernatant were collected 24hours after stimulation with the indicated cytokines. TNF-α was determined by ELISA (DuoSet, R&D Systems, Wiesbaden, Germany). Only those experiments were included into the analysis where IL-13 alone but not IFN-γ alone induced TNF-α (n=3). Journal of Investigative Dermatology 2007 127, 1271-1274DOI: (10.1038/sj.jid.5700687) Copyright © 2007 The Society for Investigative Dermatology, Inc Terms and Conditions