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J. Lee, H.-J. Moon, C.-K. Joo; Smad3 Regulate NET1, Rho GEF, in TGF-β Induced Human RPE. Invest. Ophthalmol. Vis. Sci. 2010;51(13):5965.
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© ARVO (1962-2015); The Authors (2016-present)
In previous studies we have demonstrated that RhoA dependent signaling regulate TGF-β1 induced cytoskeleton reorganization in human retinal pigment epithelium cell line, ARPE-19 cells. In addition to RhoA signaling, Rho GEF, NET1 have also been shown to mediate actin of TGF-β1. The purpose of this study were to examine what regulate Rho GEF activity and to test whether Smad signaling cross-talks with Rho pathways during actin rearrangement induced by TGF-β1.
Serum-starved ARPE-19 cells were incubated with vehicle alone or 10ng/ml TGF-β1. Using dominant negative Smad3 and active Smad3 DNA construct, we show that these proteins are critical to TGF-β1 induced cytoskeleton reorganization and NET1 expression. Actin reorganization was examined by immunochemistry and confocal microscopy. Protein expression was analyzed by Western blot analysis.
Using DNA construct targeting for Smad, we show that Smad3 is critical to TGF-β1 induced cytoskeleton reorganization and NET1, the GEF of RhoA, expression. In ARPE-19 cells that lack Smad3, TGF-β1 induced stress fiber was not observed. Interestingly, dominant negative smad3 expressing cells, TGF-β1 failed to induce the NET1 mRNA and protein expression.
we demonstrate that Smad3 regulate RhoA activation and cytoskeleton reorganization via controlling NET1 in TGF-β1 induced ARPE-19 cells. These data define a new role for Smad as a modulator of RhoA activation while regulating TGF-β1 induced epithelial-mesenchymal transitions.
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