Variable expression of cystatin C in cultured trans-differentiating rat hepatic stellate cells

Figure 3 Effect of rhCysC on TGF-β1-induced reporter gene activity and Smad2/3 phosphorylation. A: Cultured HSCs were infected with 50 MOI Ad-(CAGA)₉-MLP-luciferase followed by 1 h incubation with the indicated doses of rhCysC prior to exposure with TGF-β1 (1 ng/mL) for 4 h or left untreated. The mean ± SD of the measured luciferase activities (n = 4) was given; B: phosphorylated Smad2 (pSmad2) and Smad3 (pSmad3) were detected by immunoblotting of whole cell protein lysates prepared after treatment of HSCs with or without 500 ng/mL rhCysC for 1 h followed by incubation with 1 or 5 ng/mL TGF-β1, respectively for an additional hour. The amount of total Smad2/3 or Smad3 was used as internal loading control.