Increase of TSG-6-encoding mRNA and protein in challenged RHE (AD-like RHE model). RHEs were treated with MβCD and IL-4, IL-13, and IL-25 for 48 hours. (a) HE staining and (b) HA detection. Bar = 20 μm. (c) TSG-6-encoding mRNA expression of AD-like RHE assessed by RT-qPCR (mean ± SD; n = 3; Student t-test; ∗∗P < 0.01). (d) TSG-6 protein concentration of challenged RHE assayed using ELISA (mean ± SD; n = 3; Student t-test; ∗P < 0.05). (e) TSG-6-encoding mRNA expression of RHE treated for 48 hours by ILs or MβCD separately assessed by RT-qPCR (mean ± SD; n = 3; one-way ANOVA; ∗P < 0.05; ∗∗P < 0.01 vs. normal RHE). (f) TSG-6 and PPIB (CTL+) mRNA detection using in situ hybridization and hemalun counterstaining of RHE challenged or not challenged with IL-4 and IL-13. AD, atopic dermatitis; HA, hyaluronan; HE, hemalun/eosin; MβCD, methyl-β-cyclodextrin; RHE, reconstructed human epidermis.