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. 2011 Jan;178(1):119–127. doi: 10.1016/j.ajpath.2010.11.038

Figure 2.

Figure 2

Representative immunofluorescent confocal photomicrographs of a wounded VIC monolayer 24 hours after wounding at the WE stained for pSmad2/3 (green) (A, D, G, and J) and nuclear localization with propidium iodide (blue) (B, E, H, and K), and AWM strip parallel to WE stained for pSmad2/3 (C, F, I, and L) in wounded cultures treated with neutralizing antibody to TGF-β (15 μg/ml) (A–C), with neutralizing antibody to TGF-β (15 μg/ml) and exogenous FGF-2 (10 ng/ml) (D–F), with neutralizing antibody to FGF-2 (10 μg/ml) (G–I) and with neutralizing antibody to FGF-2 (10 μg/ml) and exogenous FGF-2 (10 ng/ml) (J–L). Same nuclear pSmad2/3 staining at the WE is observed for VICs treated with neutralizing antibody to TGF-β only and VICs treated with neutralizing antibody and exogenous FGF-2. The same nuclear pSmad2/3 staining at the WE is observed for VICs treated with neutralizing antibody to FGF-2 only and VICs treated with neutralizing antibody to FGF-2 and exogenous FGF-2. Treatment with neutralizing antibody to FGF-2 shows greater nuclear pSmad2/3 staining at the WE than VICs treated with neutralizing antibody to TGF-β at the WE. No difference was observed in pSmad2/3 staining at the AWM among the treatments. Arrows indicate direction of wound closure. M: Percentage of VICs showing nuclear pSmad2/3 staining at the WE with treatments of neutralizing antibody to TGF-β, exogenous FGF-2, and neutralizing antibody to TGF-β, neutralizing antibody to FGF-2, and exogenous FGF-2 and neutralizing antibody to FGF-2. Treatment of exogenous FGF-2 in combination with neutralizing antibody to TGF-β does not yield a statistically significant difference of pSmad2/3 staining at the WE from when treated with neutralizing antibody to TGF-β alone. Treatment of exogenous FGF-2 in combination with neutralizing antibody to FGF-2 does not yield a statistically significant difference of pSmad2/3 staining at the WE from when treated with neutralizing antibody to FGF-2 alone. A statistically significant increase is observed in percentage of VICs showing nuclear pSmad2/3 staining at the WE treated with neutralizing antibody to FGF-2 compared with WE treated with neutralizing antibody to TGF-β. Statistical significance between indicated groups at *P < 0.05, (n = 9). Original magnifications, ×400.