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Figure 6 | Respiratory Research

Figure 6

From: Proteinase-activated receptor 4 stimulation-induced epithelial-mesenchymal transition in alveolar epithelial cells

Figure 6

PAR 4 agonist-induced EGFR phosphorylation in A549 cells. A549 cells were pretreated with or without 300 nM PP2 for 30 min, and then stimulated with AYPGKF-NH2 (100 μM) for 30–60 min followed by immunoblotting with specific antibodies for the Y1173 phosphorylated EGFR and total EGFR protein. Control; without drug treatment. (A) Representative time course of EGFR phosphorylation induced by thrombin (1 U/ml) or AYPGKF-NH2 (100 μM). As a positive control, cells were treated with EGF (10 ng/ml) for 30 min. (B) Representative Western blot after treatment with or without (control) various agents (AYPGKF-NH2, AYPGKF-NH2+PP2, 100 μM FKGPYA-NH2 or EGF) for 30 min. (C) Summarized densitometric data. Protein concentrations used for separation was 20 μg/lane (thrombin or AYPGKF-NH2 or FKGPYA-NH2 treated sample) or 10 μg/lane (EGF treated sample). Each bar represents the mean ± S.E.M. for 4–6 preparations. *P < 0.05 compared with AYPGKF-NH2 alone.

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