CFs were cultured as described previously [32 (link)–35 ]. CFs were then divided into four groups according to cell treatment, namely: (1) control group, (1%FBS-DMEM); (2) TGF-β1 treatment group (1%FBS-DMEM + TGF-β1[10 ng/ml, R&D Systems]); (3) IL-25 treatment group(1%FBS-DMEM + IL-25 [10 ng/ml, R&D Systems]); and (4) IL-25 + PI3K-antagonist group (1%FBS-DMEM + IL-25 [10 ng/ml] + LY294002 [15 μM, Sigma-Aldrich)]). Cells were then cultured for another 24 h and then harvested for further analyses, including real-time PCR, western blotting, ELISA, and cell proliferation and migration analyses.
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