Unless stated otherwise, cells were seeded into six-well plates for 24 hours (murine fibroblasts at 5 × 105 cells per well; human fibroblasts at 7 × 105 cells per well) and changed into medium with 0.1% FBS (murine fibroblasts) or without FBS (human fibroblast cells) for 24 hours, and reagents were added for the indicated times. Cellular lysates were prepared on ice with modified radioimmunoprecipitation assay (RIPA) buffer [50 mM tris (pH 7.4), 1% Triton X-100, 0.25% sodium deoxycholate, 150 mM NaCl, 1 mM EDTA (pH 8), and 10 mM NaF] containing cOmplete Protease Inhibitor (Roche). After removal of insoluble material (centrifugation at 16,200g at 4°C for 15 min), proteins were separated by electrophoresis on 10% SDS–polyacrylamide gel electrophoresis and processed for Western analysis. Detection was performed using enhanced chemiluminesence (Thermo Fisher Scientific). Commercial antibodies are provided in table S2. Rabbit anti-pSmad3 antibody was used at 1:5000 dilution and generated to the peptide COOH-GSPSIRCSpSVpS (79).

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