Fig. 3 | Mucosal Immunology

Fig. 3

From: The IFN-γ–p38, ERK kinase axis exacerbates neutrophilic chronic rhinosinusitis by inducing the epithelial-to-mesenchymal transition

Fig. 3

p38, ERK signaling is crucial for IFN-γ-induced EMT in hNECs and is activated in patients with neutrophilic NE-NP. a hNECs treated with IFN-γ (100 ng/ml) in a time-dependent manner were immunoblotted using the indicated antibodies. b Western blot analyses of cytoplasmic (C) and nuclear (N) fractions from confluent hNECs, which had been treated with or without of IFN-γ. c STAT1-knockdown hNECs were treated with IFN-γ for 48 h. Immunoblots were performed on cell lysates with indicated antibodies. d Real-time qRT-PCR and western blot to analyze expression of ICSBP for the indicated times after treatment of IFN-γ. e ICSBP-knockdown hNECs were treated with IFN-γ for 48 h. Cell lysates were immunoblotted with the indicated antibodies. f Immunoblot analysis in hNECs treated with IFN-γ for the indicated times. g ICSBP-knockdown hNECs were cultured with IFN-γ for 48 h. Activation of p38, ERK was evaluated by using western blot. h hNECs were pre-treated with p38 inhibitor (SB203580), ERK inhibitor (PD98059) respectively or both followed by IFN-γ stimulation for 8 h. Cell lysates were immunoblotted with indicated antibodies. i Immunohistochemical staining of phospho-p38 and phospho-ERK in UP mucosa from control patients without nasal disease (n = 7), UPs from patients with CRSsNP (n = 11), and UP (n = 9) and NP tissues (n = 18) from patients with CRSwNP. Scale bars, 50 µm. (j, k) Comparison of nuclear p38, ERK expression in the same patients as indicated above. In all immunoblot analysis (a–h), protein intensities were quantified by ImageJ software (n = 1, three independent experiments). Representative western blots are depicted and expressed as relative band intensity adjusted to the highest band intensity of each protein. The data are representative of three independent experiments with similar results. Statistical significance was determined by Student’s t-test (d) and Kruskal–Wallis tests (P < 0.01), followed by Mann–Whitney U-test for pairwise comparisons (j, k). *P < 0.05; **P < 0.01. Data are shown as means ± s.e.m

Back to article page