Fig 2: TSLP is important for ILC2-independent Th2 cell response(A–I) OVA/alum-induced airway inflammation model was used as described in Figure 1I. The total cell numbers of eosinophil and Th2 cells were measured by flow cytometry, and the values were plotted (mean ± SEM; n = 4–12; **p < 0.01, ***p < 0.001, ****p < 0.0001, Student’s t test).(J) A modified experimental procedure for OVA-induced pulmonary allergy model for testing the effect of TSLP administration.(K and L) The total cell numbers of Th2 cells in mice treated in (J) were measured by flow cytometry, and the values were plotted (mean ± SEM; n = 5–6; *p < 0.05, **p < 0.01, Student’s t test).Results are representative of two (G–I and K–L) to three (A–F) independent experiments. See also Figure S4.

Fig 3: The production of TSLP by keratinocytes was increased in Stat3 cKO mice, which recruited more target cells in AD-like skin inflammation. (A) Analysis of TSLP expression level (pg/ml) in the ear skin of Stat3f/f mice and Stat3 cKO mice after DNCB or vehicle induction by ELISA (n = 3-4). (B) IF staining showed that TSLP was primarily expressed in keratinocytes (red). Bar = 50 μm. (C) IF staining revealed that the infiltration of TSLPR+ cells (green), CD11c+ cells (red), and CD68+ cells (red) was increased in the dermis of DNCB-treated Stat3 cKO mice compared with Stat3f/f mice. (D) Statistical analysis of the abovementioned cells (n = 3-4). The data are presented as the mean ± SD; P > 0.05 denotes ns; *P < 0.05, **P < 0.01, and ***P < 0.001.

Fig 4: DNCB-induced AD-like skin inflammation of Stat3 cKO mice was alleviated by an anti-TSLP neutralizing antibody. (A) Representative photographs of DNCB-treated Stat3f/f mice and Stat3 cKO mice after anti-TSLP antibody or isotype IgG antibody injection on Day 8. (B) The ear thickness (mm) and dermatitis score were measured every other day (n = 5-8). (C) Analysis of the histological changes in ear skin by HE staining (Bar = 100 μm), mast cells by toluidine blue staining (red arrows, Bar = 50 μm), and TSLPR-positive cells by IF staining (green, Bar = 50 μm). The data are presented as the mean ± SD; P > 0.05 denotes ns; *P < 0.05, **P < 0.01, and ***P < 0.001.