Fig. 6

TAF decreased the expression level of phosphorylated-Smad3 and down-regulated α-SMA and collagen 3α1 by up-regulating NS5ATP9 expression in activated HFL1 cells. a Protein expression of phosphorylated-Smad3 and Smad3 under different intervention conditions. Cells were serum-starved overnight, before incubation with TGF-β1 (5 ng/ml) for 24 h, and then treated with SD-208 (1 μmol/L) or TAF (250 μmol/L) for 24 h. Phosphorylated and total Smad3 were measured by Western blotting against phosphorylated-Smad3 and Smad3. Blots were analyzed by densitometry and the results were expressed as relative units. All data were demonstrated as fold changes of gene expression normalized to β-actin. b Protein expression of α-SMA, collagen 3α1 and NS5ATP9 in cells stimulated by TGF-β1. Cells were serum-starved overnight before activated with TGF-β1 (5 ng/ml) for 24 h. Finally, cells were collected and lysed, and used for protein levels. All the objectives were determined by Western blotting. Blots of collagen 3α1, α-SMA and NS5ATP9 were analyzed by densitometry, and the results were expressed as relative units. Data were presented as mean ± standard error of the mean (SEM) of three independent trials performed in duplicate, and statistical significance across intervention groups was determined using one-way ANOVA. #P < 0.05 vs. untreated cells. *P < 0.05 vs. control (TGF-β1-treated) cells