Fig. 2

Pam3 CM up-regulates fibrogenic and osteogenic mediators in AVICs independent of TLR2. A–C AVICs were exposed to Pam3 (0.03 μg/ml), control CM or Pam3 CM for 48 h. Levels of collagen I, collagen IV, MMP-2, MMP-9, ALP and RUNX2 were determined by immunoblotting. Representative immunoblots and densitometric data show that Pam3 CM selectively up-regulated collagen I, MMP-2 and ALP in AVICs among the fibrogenic and osteogenic factors examined while control CM or a low concentration of Pam3 had no effect. D AVICs were pretreated with TLR2 inhibitor CU CPT 22 (10 µM) or DMSO for 1 h and then exposed to Pam3 CM treatment for 48 h. Inhibition of TLR2 in AVICs did not alter the effect of Pam3 CM on the upregulation 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