, Blots were blocked with Tris Buffered Saline (TBS) (50 mM Tris, 150 mM NaCl) containing 5% low-fat dried milk/0.1% Tween 20 (for the anti-glyceraldehyde 3-phosphate dehydrogenase [GAPDH] antibody) or 5% bovine serum albumin (BSA)/0.1% Tween 20. Membranes were probed overnight at 4 °C with either anti-GAPDH (1:20,000; G9545 Sigma-Aldrich), Cell Signaling Technology) antibodies in corresponding blocking solutions. After washing, membranes were incubated with a goat anti-rabbit horseradish peroxidase-conjugated secondary antibody, vol.55
, Intensity of the single band corresponding to SMAD3 or to its phosphorylated form p-SMAD3 was normalised with the intensity of the GAPDH band, before the p-SMAD3/SMAD3 ratio was calculated, ChemiDoc TM MP Imaging System (Biorad)
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