Figure 5. Effect of recombinant transforming growth factor (TGF)-β2 on localization of Smad2/3 with Co-Smad4 in lamina cribrosa (LC) cells. LC cells were incubated with recombinant TGF-β2 (5 ng/ml) for 60 min. Cells were then fixed with 3.5% formaldehyde solution and were stained with antibodies for pSmad3, pSmad2, and Smad4. A: Co-localization of pSmad3 (red) and Smad4 (green) in untreated LC cells, and B: in TGF-β2 treated cells. C: Co-localization of pSmad2 (red) and Smad4 (green) in untreated LC cells, and D: in TGF-β2 treated cells. E: Negative controls consisted of LC cells incubated with rabbit and mouse IgG and were co-stained with nuclear stain DAPI (blue).