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Fig. S1 Skin phenotype of 2-month-old K14-TSLP mice. (A) Immunostaining of epidermal sheets showing the expression of TARClCCL17 and MDClCCL22. (B) Expression of maturation markers at the cell surface of LC analysed by flow cytometry. Histograms demonstrate the overlay of CD40, CD86 and CCR7 expression on LC, i.e. on cells gated on MHC-class II. The plain black Histogram designates cells isolated from K14-TSLP mice, the grey line cells isolated from littermate control mice and the dotted grey line the isotype control. (C) Expression of maturation markers at the cell surface of dermal DC analysed by flow cytometry. Cells were gated on MCH-class II+Langerin- cells. One representative picture is shown for immunofluorescence stainings or flow cytometry analyses. Groups of three mice were used.

Fig. S2 Phenotype of dermal langerin+CDIIc+MHCclass II+ cells. Expression of maturation markers at the cell surface of dermal langerin+ DC were analysed by flow cytometry. Cells from mouse ears treated with vehicle (ethanol, ETOH) or MC903 for 4 days (nght panel) or from 2-month-old K14-TSLP transgenic (K14-TSLP) mice or littermate control (control) mice (left panel) were used, n 3, 4 Dot plots show cells gated on MHC-class II and CDIIc and one representative image of the results.

Fig. S3 0x40-LiCD134 expression and cytokine production by lymph node DC in early AD i.e. before the development of inflammation: Mice were treated with vehicle (ethanol, ETOH) or MC903 for 4 days on ears Oxazolone-treated mice were used as a control for a Thl predominant response. Dot plots show cells treated on CDIIc and one representative image of the results. Numbers show relative percentage of cytokine+ or 0x40-L+ langerin+ DC and of cytokine+ or 0x40-L+ langerin- DC, gated on CDIIc.

Fig. S4 Phenotype of CD4+ T cells In K14–TSLP mice: Production of IFN-g and IL-13 by lymph node CD4+ T cells analyzed by flow cytometry in K14-TSLP transgenic (K14-TSLP) and littermate control (WT) mice, n ∇ 2–4. Oxazolone-treated mice were used as a positive control for a Thl predominant response, n ∇ 3,4. Data were analysed using a student’s t-test.

Fig. S5 Phenotype of three-month old K14-TSLP mice: (A) Expression of maturation markers at the cell surface of LC analysed by flow cytometry, n 3. (B) Expression of maturation markers at the cell surface of dermal DC analysed by flow cytometry. Results from 4 animals were pooled. (C) Number of emigrated DC analysed in the lymph nodes by flow cytometry, n = 4.

Fig. S6 Depletion of langerin+ skin DC: (A) Immunohistochemistry of epidermal sheets showing the expression of Langerin (red fluorescence) and MHCclass II (green fluorescence) in the epidermis of wild type (WT) and LC-depleted (DTR-LC) mice treated with vehicle (ethanol, ETOH) or MC903. (B) Flow cytometry analysis of langerin expression in the dermis of wild type and LC-depleted mice treated with vehicle (ethanol) or MC903. Dermal cells were gated on MHC-class I1. Representative images are shown. Groups of three to eight animals were used.

Fig. S7 Quantitative PCR for TSLP transcript in mouse ears. Wild type (WT) and LCdepleted (DTR-LC) mice were injected with diphthena toxin (DT) at day-2, day+4 and day+10 of the treatment. DTR-LC mice not injected with diphtheria toxin were used as additional controls. Ethanol (vehicle. ETOH) and MC903 were topically applied to mouse ears for 15 days. Then RNA was prepared from whole ears Groups of three to seven animals were used.

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Please note: Wiley Blackwell is not responsible for the content or functionality of any supporting information supplied by the authors. Any queries (other than missing content) should be directed to the corresponding author for the article.