Figure 5.

IL-27 stimulation does not affect TGF-β-mediated activation of Smad proteins in LX2 cells and rat HSC. A, B: LX-2 cells or rat HSC were pretreated with IL-27 (30 ng/mL) or IFNγ (30 ng/mL) for 6 h (A) or 12 h (B). Expression of Smad2 (A) or phosphorylation and expression of Smad3 (B) was monitored by Western blot analysis. Staining for tubulin, pY-STAT1 and STAT1 are provided as controls. C, D: LX-2 cells or rat HSC were pretreated with IL-27 (30 ng/mL) or IFNγ (30 ng/mL) for 6 h (C) or 12 h (D) and were subsequently stimulated with TGF-β (1 ng/mL) for 0.5 h. Expression of Smad2 (C) or phosphorylation and expression of Smad3 (D) was monitored by Western blot analysis. Staining for tubulin, pY-STAT1 and STAT1 are provided as controls. E: Induction of a Smad2/Smad3/Smad4-dependent CAGA reporter construct after treatment of LX-2 cells with IL-27 (30 ng/mL), TGF-β (1 ng/mL) or both for 2 h or 16 h. Standard deviations for a triplicate experiment are given as error bars.