Figure 5
Recombinant soluble CD109 protein inhibits transforming growth factor beta 1-induced extracellular matrix and CCN2 production. (A) Systemic sclerosis (SSc) and normal skin fibroblasts were left untreated (-) or treated (+) for 24 hours with 100 pM transforming growth factor beta 1 (TGF-β1) in the presence of 1.0 nM recombinant CD109 protein or PBS/0.1% BSA vehicle control. Cell lysates were prepared and analyzed by western blot using anti-fibronectin (top panel), anti-type I collagen (second panel) or anti-CCN2 (third panel) antibodies. Membranes were reprobed with an anti-β-actin (bottom panel) antibody to confirm that equal amounts of protein were loaded in each lane. Results presented are representative of three independent experiments. (B) Densitometric analysis of the data from the above blot was performed using β-actin as a loading control. The representative SSc fibroblast data shown correspond to (A) SSc Patient S21 and (B) SSc Patients S19, S2 and S16.