Fig. 2

Paracrine TGF-β1 was enssential for CAFs induced EMT. a The amount of TGF-β1 released into the cell culture supernatant of cells treated with CAF-CM was determined by ELISA. b The wound-healing assay of MCF-7 and MDA-MB-231 cells treated with recombinant TGF-β1, CAF-CM, the TGF-β1 inhibitor SB431542 (SB) or pirfenidone (PFD), respectively. c SB or PFD treatment impaired the CAF-mediated increase in invading cancer cells, as indicated by the Transwell assay. d Western blotting analysis for E-cadherin, vimentin and β-catenin of cells treated as in (b). e Immunofluorescence staining showed that treatment with SB or PFD increased E-cadherin levels, decreased vimentin and nuclear β-catenin distribution in MCF-7 and MDA-MB-231 cells. CAFs promoted F-actin polymerization and stability in cancer cells, whereas treatment with SB or PFD impaired the formation of stress fibers. Scale bar: 15 μm