Figure 4: Effects of TSG-6 on ECM Expression in Human VSMCs

(A) HASMCs were incubated for 24 h with the indicated concentrations of TSG-6 and were harvested for immunoblotting analyses of collagen-1, collagen-3, TIMP-2, MMP-2, MMP-9, fibronectin, elastin, and α-tubulin. (Top) Representative results of protein expression of each molecule. (Bottom) Densitometric data of each molecule after normalization relative to α-tubulin. n = 4 to 7; *p = 0.025; †p = 0.015; ‡p = 0.047; #p < 0.01 vs. 0 ng/ml of TSG-6. (B) HASMCs were incubated for 24 h in serum-free conditioned medium with the indicated concentrations of TSG-6. In the culture supernatant, MMP-2 and -9 activities were determined by the gelatin zymography. n = 5 to 6; §p = 0.049; ¶p = 0.031; @p = 0.011; &p < 0.0001 vs. 0 ng/ml of TSG-6. All values are presented as mean ± SEM. ECM = extracellular matrix; M = matrix metalloproteinase marker; MMP = matrix metalloproteinase; S = sample; TIMP = tissue inhibitor of metalloproteinase; VSMC = vascular smooth muscle cell; other abbreviations as in Figure 1.