Fig. 5

OSM suppresses SMAD signaling. a After pretreatment with the OSM (10 ng per ml) or IL6 (20 ng per ml), C3H/10T1/2 cells were stimulated with TGF-β1 (2.5 ng per ml, 12 h) and the relative expression level of αSMA mRNA was calculated. The one-way ANOVA and Dunnett’s multiple comparisons test were used for the statistical analysis (F (2, 6) = 97.35). b Immunoblot analysis of phospho-ERK1/2, total ERK1/2, phospho-SMAD2 linker (Ser245/250/255) and total SMAD2 protein. Thirty minutes after U0126 (20 nM) pretreatment, C3H/10T1/2 cells were stimulated with OSM (10 ng per ml) or IL6 (20 ng per ml) for 30 min. and collected for the analysis. U0126: a selective MEK1/2 inhibitor. c Immunoblot analysis of phospho-SMAD2 C-terminal (Ser465/467) and Lamin A/C. Thirty minutes after OSM (10 ng per ml) pretreatment, C3H/10T1/2 cells were stimulated with TGF-β1 (2.5 ng per ml, 12 h) and collected at indicated time points. d After pretreatment with U0126 (20 μM, 60 min) and OSM (10 ng per ml, 30 min), C3H/10T1/2 cells were stimulated with TGF-β1 (2.5 ng per ml, 12 h) and the relative expression level of αSMA mRNA was calculated. Two-tailed t-test with Welch’s correction was used for the statistical analysis (t = 3.212, df = 3.172). Data show the mean and the standard deviation (error bar) of technical triplicates from a representative experiment (a, d). *p < 0.05