Figure 8 of Zode, Mol Vis 2011; 17:1745-1758.


Figure 8. Effect of Smad2 or Smad3 siRNA on transforming growth factor (TGF)-β2 stimulation of fibronectin (FN) and plasminogen activator inhibitor (PAI)-1 in optic nerve head (ONH) astrocytes and lamina cribrosa (LC) cells. A: ONH astrocytes and LC cells were treated with small interfering RNA (siRNA) controls (lane 1=Non-targeting siRNA; lane 2=RISC-Free siRNA) or with Smad3 siRNA (25 nM and 50 nM; lanes 4–5) for 48 h, and were then treated with recombinant TGF-β2 (5 ng/ml) for 24 h. Cellular FN, PAI-1, total Smad3, and actin were assessed by western blot. B: Relative densities of Smad3 and actin in ONH astrocytes were measured using densitometric analysis of the western blots. Smad3 was normalized to actin and the fold change in Smad3 over the vehicle control was plotted (n=3,* p<0.001 versus control). C and D: Relative FN or PAI-1 was normalized to actin and the fold change in FN (C) or PAI-1(D) over the vehicle control was plotted (n=3,* p<0.001 verses TGF-β2 treated). E: ONH astrocytes and LC cells were treated with siRNA controls (lane 1=Non-targeting siRNA; lane 2=RISC-Free siRNA) or with Smad2 siRNA (lanes 4–5, 100 nM in duplicate) for 48 h, and were then treated with recombinant TGF-β2 (5 ng/ml) for 24 h. F: Cellular FN, PAI-1, total Smad2, and actin were assessed by western blot. Smad2 levels in ONH astrocytes were normalized to actin and the fold change in Smad2 over the vehicle control was plotted (n=3, *p<0.0018 verses control). G and H: Relative densities of FN, PAI-1, and actin were measured using densitometric analysis. Relative densities of FN or PAI-1 were normalized to actin and the fold change in FN (G) or PAI-1(H) over vehicle the control was plotted (n=3, p<0.0048 for FN and p<0.0015 for PAI-1, * verses TGF-β2 treated).