Effect of transforming growth factor-β1 on human intrahepatic cholangiocarcinoma cell growth

Figure 2 TGF-β1 stimulates the TGF-β signaling pathway. A: The addition of 0.1 and 1 ng/mL TGF-β1 augmented Smad2 phosphorylation (phospho-Smad2), p15^Ink4B, and PAI-1 proteins in Western blotting of HuCCT1 cells; B: TGF-β1 stimulation for 48 h enhanced p15^Ink4B mRNA expression in HuCCT1 cells in RT-PCR: ^aP < 0.05 vs absence of TGF-β1; C: TGF-β1 stimulation for 48 h also augmented PAI-1 mRNA expression in HuCCT1 cells in RT-PCR: ^aP < 0.05 vs absence of TGF-β1; D: PAI-1 mRNA was determined by RT-PCR after incubating the HuCCT1 cells with 5 ng/mL neutralizing anti-TGF-β1 antibody or nonimmune control IgG for 96 h. The addition of anti-TGF-β1 antibody attenuated the level of PAI-1 mRNA: ^aP < 0.05 vs control IgG; E: The addition of TGF-β1 to HuCCT1 for 6 h led to a concentration-dependent increase in TGF-β1 mRNA expression by RT-PCR: ^aP < 0.05 vs absence of TGF-β1; F: TGF-β1 mRNA in HuCCT1 was determined by RT-PCR after incubating the cells with 5 ng/mL neutralizing anti-TGF-β1 antibody or nonimmune control IgG for 96 h. The addition of anti-TGF-β1 antibody attenuated the level of TGF-β1 mRNA: ^aP < 0.05 vs control IgG.