Fig 2: TSLP promotes CD4+ T cell proliferation and Th17 cytokine expression. (A) CD4+ T cells were stained with CFSE and then cultured with or without 100 ng/mL rmTSLP for 4 days, flow cytometry was performed to detect the proliferation of CD4+ T cells. (B) Quantification of cell proliferation rate in (A). CD4+ T cells were cultured with rmTSLP (100 ng/mL) for 1, 2, and 3 days. (C) qRT-PCR and (D) ELISA were performed to detect the mRNA and protein levels of IL-17A, IL-17F, and IL-22 in CD4+ T cells. (E) Flow cytometry was conducted to analyze the protein levels of IL-17A in CD4+ T cells. (F) Quantification of IL-17A levels in (E). Ctr, control. (Data are mean ± SEM, *P < 0.05, **P < 0.01, n = 3).

Fig 3: Imiquimod increases IL-17 and IL-22 in both ?d and aß T cells in skin.Skin and ear of wild type C57BL/6 mice were treated by topical application of imiquimod cream (Fougera, n=4) or control cream (Vaseline, n=3) for 6 consecutive days. (a) Weight loss of imiquimod and control cream treated mice monitored daily. (b) Photographs of imiquimod and control cream treated skin and ear; photos were taken at day 6. H&E stained ear and back skin sections of imiquimod treated and control mice. Scale bar, 200 µm. (c) Thickness of skin measured by Digimatic Caliper at day 6 in control and imiquimod treated mice. Data showed represents average of at least two measurements. (d) Ear thickness of imiquimod and control cream treated mice monitored daily. Ear thickness was measured using Digimatic Caliper. (e) Quantitative PCR analysis of Th17 associated cytokines and Foxp3 in skin after 6 days of imiquimod and control cream treatment. (f) Representative flow cytometric analysis of TCR?d+V?4+IL-17+ (upper raw) and TCR?d+V?4+IL-22+ cells (lower row) in the skin of control (n=3) and imiquimod treated mice (n=4). (g) Representative flow cytometric analysis of single-positive TCRß+IL-17+ (upper raw) or TCRß+IL-22+ (lower raw) cells in the skin of control (n=3) and imiquimod treated mice (n=4). Data representative of more than three experiments, results are shown as mean±s.e.m., significance determined by unpaired two-tailed Student's t-test (*P<0.05; **P<0.01; ***P<0.001).

Fig 4: Aloperine inhibits the migration and Th17-inducing effect of DCs. (A) A graphical presentation of the air pouch model. On the first day, 3 ml of sterile air was injected subcutaneously into a shaved skin site on the back of each mice. Then, the pouches were settled for 3 days to heal the wound. On the third day, this action was repeated. On the sixth day, LPS was injected into the pouches with or without aloperine. The exudates were collected for subsequent analysis 24 h later. (B) Quantitative analysis of total cell numbers in the skin. (C–E) Representative flow cytometry analysis of neutrophils (CD11b+MHC-II-Ly6G+Ly6C-), macrophages (CD11b+F4/80+), and DCs (CD11c+F4/80-). (F) Western-blot analysis for p-P65 and ß-actin. (G) Real-time PCR analysis of IL-17A, IL-23, and ROR?t in CD4+ T-cells receiving DC cultural supernatant pretreated by LPS + PBS or LPS + aloperine. The values are presented as the mean ± SEM (n = 5 mice per group). *p < 0.05; **p < 0.01; and ***p < 0.001.

Fig 5: Th1, Th17 and IL-17A-producing ?dT17 cells be detected in K14-WFDC12 transgenic mice and WT mice after IMQ-induced psoriasis model. (A) Representative of Th1 cells secreting IFN-? and IL-17-producing cells (Th17 cells) in the lymph nodes of mice after IMQ-induced. (B, C) Quantification of Th1 cells and Th17 cells in the lymph nodes of mice (proportion of CD4+ cells Gate), n = 5/group. (D, E) IFN-? and IL-17A mRNA expression level of psoriasis-like lesion areas in K14-WFDC12 transgenic mice and WT mice. (F, G) Representative FCM images and quantification indicated percentages of IL-17A–producing ?dT17 cells in lymph nodes of K14-WFDC12 transgenic mice and WT mice treated with IMQ, n = 5/group. (H) ELISA assessed the levels of IL-17A in serum of K14-WFDC12 mice and WT mice treatment with IMQ, n = 5/group. All the data are shown in this and the figures below came from samples from individual mice unless stated otherwise. The data are presented as M ± SD. **p < 0.01. ns, not significance. (unpaired Student’s t-test).