Fig. 3.

RANTES contributes to the up-regulation of fibrogenic mediators, and TNF-α mediates Pam3 CM-induced up-regulation of both fibrogenic and osteogenic mediators. A ELISA assay found that Pam3 CM has greater levels of TNF-α and RANTES in comparison to control CM. Data are presented as mean ± SEM. n = 5, *P < 0.05 vs. control CM. B and C AVICs were treated for 48 h with Pam3 CM pre-incubated with neutralizing antibodies (10 μg/ml) against RANTES or TNF-α. Non-immune mouse IgG (10 μg/ml) was added to Pam3 CM as a control. Neutralization of RANTES reduced the potency of Pam3 CM on the expression of collagen I and MMP-2 while neutralization of TNF-α attenuated the effect of Pam3 CM on the levels of collagen I, MMP-2 and ALP. Data are presented as mean ± SEM. n = 5 cell isolates from distinct donor valves in each group. *P < 0.05 vs. untreated control. ^#P < 0.05 vs. Pam3 CM and Pam3 CM with IgG