Murine Model of Allergic Skin Inflammation

PCA was induced as follows^{82 (link)}: briefly, both ears of mice were intradermally injected with 10 ug/ml IgE (25ul, D8406, Sigma-Aldrich). The tail vein, 16 h later, was injected with 1 mg/ml DNP-HSA (100ul, D-5059, Biosearch Technologies). After 30 min, a full thickness biopsy of the right ear was fixed and embedded for toluidine blue staining. Evans blue was extracted from the remaining skin tissue of the right ear at 65 °C for 16 h, and its absorbance was measured at 610 nm. After 12 h, blood was collected, and the concentration of IL4, IL13, and IL10 in plasma/ear tissue was determined by ELISA according to the manufacturer’s instructions (Jianglai Biotechnology Co., Ltd., China). The concentration of LPS in plasma and histamine in plasma/ear tissue was determined by ELISA (USCN Life Science and Technology, China). The left ear tissue was frozen in liquid nitrogen and stored at −80 °C for subsequent qPCR detection.