Fig. 2

Evaluation of the protein synthesis of myofibroblast phenotype markers and extracellular matrix macromolecules in cultured human systemic sclerosis (SSc) skin fibroblasts. Western blotting and related densitometric analysis of the protein synthesis of α-smooth muscle actin (α-SMA), S100A4, type I collagen (COL-1) and fibronectin (FN) in cultured human SSc skin fibroblasts maintained in normal growth medium (untreated), treated with selexipag at the concentration of 30 μM, 3 μM, and 0.3 μM, and treated with ACT-333679 at the concentration of 10 μM, 1 μM, and 0.1 μM, for 48 h. For each experimental condition, the value of the synthesis of α-SMA, S100A4, COL-1, and FN was normalized to that of the corresponding glyceraldehyde 3-phosphate dehydrogenase (GAPDH). The resulting value with each treatment was compared to that of the untreated cells (taken as a unit value), in order to obtain the level of protein synthesis. A molecular weight (MW) lane was also included. The final results represent the mean ± standard deviation (SD) of the values obtained from six independent experiments on cultured human SSc skin fibroblasts: *p < 0.05 and **p < 0.01 vs. untreated cells. kDa, kiloDalton