Figure 5
7-Methoxyisoflavone attenuated FITC-induced AD symptoms by regulating Th1/Th2 balance and MAPK-AP1 signaling pathway. (a) Heatmap of all significant different chemokines related genes between model (FITC) and drug-treated mice ear tissue. (b) Relative Cxcl9 (p = 0.0066), and Cxcl10 mRNA levels were determined by quantitative RT-PCR. (c) Relative Ccl17 (p = 0.0096) and Ccl22 (p = 0.0072) mRNA levels were determined by quantitative RT-PCR. (d) Total serum IFN-γ and IL-4 levels were measured by enzyme-linked immunosorbent assay (ELISA). (e) The ratio of IL-4 to IFN-γ was calculated (p = 0.0318) (n = 10). (f) TSLP proteins labelled with FITC fluorescent dye was shown in green. Nuclei labeled with DAPI was shown in blue. (g) The average fluorescence intensity was shown as mean ± SEM. (n = 6). (pmodel < 0.0001, two-tailed, p7-met = 0.0053, two-tailed). (h) The phosphorylation of MAPKs and expression levels of TSLP in the ear extracts was analyzed by Western blotting. (i) The phosphorylation of MAPKs and STAT3 in the dorsal skin extracts was analyzed by Western blotting. (j) The expression levels of p-IκBα, p-p65 (N) and the nuclear translocation of AP-1in the ear extracts were analyzed by Western blotting. The expression levels of Ras, c-Raf, p-ERK, ERK, p-JNK, JNK, p-p38 and p38 in the total protein and the nuclear translocation of c-Jun and c-Fos of the right ears were detected by Western blotting. FITC fluorescein isothiocyanate, 7-Met 7-Methoxyisoflavone, TSLP thymic stromal lymphopoietin, Ccl C–C motif chemokine ligand, Cxcl C-X-C motif chemokine ligand; *p < 0.05, **p < 0.01, ***p < 0.001 and ****p < 0.0001. Interleukin; IFN Interferon, FITC fluorescein isothiocyanate, OXZ oxazolone, 7-Met 7-Methoxyisoflavone, p65 (N) nucleus p65, p-ERK phospho-p44/42MAPK(ERK1/2), ERK p44/42MAP Kinase, p-p38 phospho-p38 MAPK(Thr180/Tyr182), p38 p38 MAPK, c-Fos (N) nucleus c-Fos, c-Jun (N) nucleus. In order to avoid the interference of non-specific binding of antibodies to chemiluminescence imaging. Under the premise of ensuring credibility, some WB images are cropped strips, all images have been provided in the Supplementary Document.
