Fig 2: Highly expressed Galectin‐7 (Gal‐7) in both skin lesions and sera from atopic dermatitis (AD) patients.(A) Amount of Gal‐7 in tape‐stripped stratum corneum samples in AD and healthy control (HC) measured by liquid chromatography/mass spectrometry analysis (AD, n = 8; HC, n = 3). (B) Representative immunohistochemical staining and image analysis for Gal‐7 in skin lesions from AD and HC. The boxed areas were enlarged in the left panels. Image analyses showed the intensity of Gal‐7 expression in intercellular spaces of epidermal keratinocytes (red lines, strong positive; blue lines, middle to weak positive). Scale bars = 100 μm. (C) Percentage of intercellular Gal‐7+ epidermal keratinocytes in AD and HC (AD, n = 11; HC, n = 4). (D), Serum levels of Gal‐7 were measured by ELISA in AD patients and HC (AD, n = 20; HC, n = 7). (E, F) Correlation between serum Gal‐7 levels and transepidermal water loss (TEWL) (n = 20) or SCORing AD (SCORAD) (n = 17). Data were shown as means ± SD. *P < .05; n.s., not significant

Fig 3: Galectin-7 (Gal-7) is required to avoid interleukin (IL)-4/IL-13–induced disruption of cell-to-cell and/or cell-to-extracellular matrix (ECM) adhesion junction of the epidermis. (A) Three-dimensional (D)–reconstructed epidermis transduced with short hairpin control (shCtr) or shGal-7 was treated with or without IL-4/IL-13 for 48 hours at a concentration of 50 ng/mL Gal-7, E-cadherin, Desmocollin-1 (DSC1), Claudin-1 (CLDN1) and Desmoglein-1 (DSG1) mRNA levels were assessed by real-time reverse transcriptase-PCR. (B) Representative images of haematoxylin and eosin (HE) and immunohistochemical (IHC) staining for Gal-7 in shCtr or shGal-7–transduced 3D-reconstructed epidermis treated with or without IL-4/IL-13 for 48 hours at a concentration of 50 ng/mL. The boxed areas were enlarged in the right panels. Black arrows indicate acantholysis. Scale bars = 50 µm. Data were shown as means ± SD of two or three independent experiments. *P < .05. NT, no treatment

Fig 4: Interleukin (IL)-4/IL-13–mediated Galectin-7 (Gal-7) release from normal human epidermal keratinocytes (NHEKs). (A) NHEKs were treated with or without IL-4/IL-13, interferon (IFN)-? and IL-17/IL-22 for 48 h at a concentration of 50 ng/mL Gal-7 mRNA levels were assessed by real-time reverse transcriptase-PCR. (n = 3). (B) Gal-7 protein levels in culture supernatants were measured by ELISA (n = 3). (C) IL-4/IL-13–induced cell death was analysed by flow cytometry (n = 6). (D) Confocal images of immunostaining for Gal-7 (green). Nucleus, actin filaments and tight junctions were counterstained with DAPI (blue), phalloidin (red) and ZO-1 (yellow), respectively. Scale bar = 30 µM. (E) Representative images of haematoxylin and eosin (HE) and immunohistochemical staining for Gal-7 in 3-dimensional (D)–reconstructed epidermis treated with or without IL-4/IL-13 for 48 hours at a concentration of 50 ng/mL. Scale bars = 50 µm (upper and middle) and 10 µm (bottom). Black arrows indicate intercellular deposits of Gal-7. Data were shown as means ± SD of three independent experiments performed in one (A, B) or duplicate (C). *P < .05, **P < .01; n.s., not significant