Figure 5.

ConA/HSC-induced hepatocyte injury is ameliorated by anti—IFN-β Ab and JNK inhibitor. Immunofluorescence staining (A) and Western blot analysis (B) show increased intracellular accumulation of IFN-β in ConA-treated HSCs. Bar graph shows relative densitometry values of IFN-β normalized to internal control (β-actin). C: HSCs were incubated for 8 hours in a medium containing 50 μg/mL ConA. Anti—IFN-β Ab (1 μg/mL) or JNK inhibitor (JNKi) (SP600125; 10 μg/mL) was added to the medium before transfer to hepatocytes. After 12 hours of incubation, 2′,7′-dichlorofluorescin diacetate staining was performed to determine oxidative stress. Bar graph shows mean fluorescence intensity (MFI). D: Protein lysates from hepatocytes incubated in ConA/HSC medium without or with anti—IFN-β Ab or SP600125 were subjected to Western blot analysis to detect cleaved caspase-3. Bar graph shows relative densitometric values of caspase-3 normalized to β-actin. E: HSCs were incubated for 8 hours in a medium containing 50 μg/mL ConA; medium was transferred to hepatocytes. After 12 hours of incubation, nuclear translocation of IRF1 was determined. All images are representative of three separate experiments. Bar graphs show combined values from three independent experiments. Data are expressed as means ± SD (B–E). *P < 0.05, **P < 0.01, and ***P < 0.001. Scale bar = 100 μm (A and C). Veh, vehicle.