Fig. 1.

Effect of FXa, thrombin on LX2 cells in vitro. a & b Incubation of cells with control medium alone resulting in minimal cell contraction and staining for αSMA (× 100 magnification). c & d Incubation of LX2 cells with thrombin at a dose of 10 nM resulted in some activation of LX2 cells with positive staining for αSMA and some contraction (× 100 magnification) e & f Incubation of LX2 cells with FXa at a dose of 0.5 U/ml resulted in activation of LX2 cells with positive staining for αSMA and a contracted phenotype (× 100 magnification). g & h Addition of FXa (0.5 U/ml) in combination with thrombin (10 nM) resulted in activation of LX2 cells with heavy positive staining for αSMA and a contracted phenotype (× 100 magnification). i - k LX2 cells were incubated with FXa and/or thrombin. αSMA, procollagen and TGF-beta expression were analysed by qRT-PCR. Data were normalized to the endogenous housekeeping gene GADPH and fold change differences in expression relative to control (untreated LX2) cells were calculated using the Comparative ddCT method. Data are Mean ± SEM from one of the representative experiments. Significance was denoted as *p < 0.05, **p < 0.01. Abbreviations: αSMA, alpha smooth muscle actin; FXa, Factor Xa; TGF-beta, Transforming growth factor beta; Thr, Thrombin