Research Paper Volume 12, Issue 20 pp 20024—20046

Figure 6. Stellate cells are activated and produce ECM components after GDF11 exposure. (A) Activation of the LX2 stellate cell line by GDF11 (100 ng/ml) or TGF-β (100 ng/ml, positive control) (scale= 100μm). (B) The nuclear translocation ratio of SMAD2/3 complexes after GDF11 and TGF-β treatment (n=3 per group). (C) Cell viability was assessed in LX2 cells, treated with different doses of GDF11 (25, 50, 100 ng/ml) for 48 hours, by using alamarBlue™ Cell Viability Reagent. (D) Relative mRNA expression of liver fibrosis/stellate cells activation markers in LX2 cells after exposure to GDF11 (100 ng/ml) or TGF-β (100 ng/ml). (E) Protein expression levels of liver fibrosis/stellate cells activation markers in LX2 cells after GDF11 or TGF-β exposure. Protein expression of selected effectors (pSMAD2, COL1A1, αSMA, vimentin (VIM), actin, GAPDH) were quantitatively assessed by immunoblotting in LX2 cells exposed or not for 48 h to GDF11 (50 or 100 ng/ml) or stimulated for 48h with TGF-β (100 ng/ml). Images are representative of three independent experiments. (F) Data quantification represents the means ± SD. * p<0.05; ** p<0.01; *** p<0.001 (Mann-Whitney U-test).