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. 2021 Mar 24;12:639095. doi: 10.3389/fphar.2021.639095

FIGURE 2.

FIGURE 2

Gefitinib inhibits TGF-β1-induced expression of EMT markers. (A and B) Serum-deprived A549 cells were treated with TGF-β1 (5 ng/mL) or in combination with gefitinib for 72 h. (A) Protein expression of epithelial marker (E-cadherin) and mesenchymal markers (N-cadherin, vimentin, and α-smooth muscle actin) was determined by Western blot analysis. Actin was used as a loading control. (B and C) TGF-β1-induced expression of vimentin was measured by immunocytochemistry. Nuclei were counterstained with DAPI. Scale bars = 50 µm. (D–F) Serum-deprived A549 cells were incubated with TGF-β1 (5 ng/mL) and gefitinib for 24 h. After RNA extraction and cDNA synthesis, we performed qRT-PCR to measure the expression of CDH1, CDH2, and VIM mRNA using GAPDH as an internal control. # p < 0.01 vs. control; *p < 0.05, **p < 0.01 versus the group treated with TGF-β1 only.