Fig. 2

TMF and CAT attenuate TGF-β1-induced fibrosis in A549 cells with decreasing EMT. A The morphology of A549 cells was observed by 100 × microscopy, after 48 h of treating TGF-β1 (10 ng/ml) with or without TMF (2 μM) and CAT (2 μM). B Western blot results of treating A549 cells with TGF-β1 for 24 or 48 h, together with TMF or CAT. The graph shows the relatively quantified value of western blot results if that of non-TGF-β1-treated group against GAPDH was 1. C Specific time (24 h) was selected and A549 cells were treated with TGF-β1, TMF and CAT. Changes in expression of various factors were observed. Another specific time (48 h) was selected to treat A549 cells with TGF-β1, TMF and CAT. The generation of ECM by the canonical signaling pathway, the degree of progress of non-canonical signaling pathway, and the expression of NOX2 were observed. D After treating TGF-β1 with or without TMF and CAT for 48 h, the cellular levels of α-SMA (red) and E-cadherin (E-cad, green) were observed. For each observation, a merged image with DAPI is also shown. E The EMT inhibitory effects of TMF and CAT were confirmed through the wound healing assay. Compared to the time prior to treatment, microscopic images of 72 h treatments are presented in which the scratched wounds were closed according to the cell migration. For each 48 and 72 h treatment, assuming that when the wound is completely closed means 100%, the relative degree of closure of each treatment group is presented as a graph. F It was confirmed again that TMF and CAT inhibited EMT of A549 cells, through the transwell migration assay. We counted the cells that migrated across the permeable membrane from the upper layer of cell culture during the treatment