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. 2022 Jan 20;11(3):346. doi: 10.3390/cells11030346

Figure 3.

Figure 3

NEN treatment inhibited cell migration and reversed EMT in TGF-β1-induced A549 cells and DHLF-IPF cells. A wound-healing assay was performed to evaluate the effect of NEN on cell migration. The width of the scratch was photographed (upper panels) and quantified (lower panels) at 0, 24, and 48 h post-scratching of TGF-β1-induced A549 cells (A) or at 0, 12, and 24 h post-scratching of DHLF-IPF cells (B) (magnification 40×). The mRNA levels of CDH1, VIM, ACTA2, and COL1A1 in (C) TGF-β1-induced A549 cells and (D) DHLF-IPF cells treated with or without NEN for 24 h were determined by qPCR. The results were normalized to the GAPDH level. Control or TGF-β1-induced A549 cells were treated with or without NEN (0.5 μM) for 24 h. The protein expressions of α-SMA, VIM, E-cad, Col-I, and FN were measured by an immunoblotting assay (E), and the cellular localization of α-SMA and VIM were determined by immunofluorescence staining (F) (magnification 400×, bar = 50 μm). (G) DHLF-IPF cells were exposed to NEN (1 μM) for 24 h, and the protein expressions of α-SMA, VIM, and FN were detected by an immunoblotting assay. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.