Figure 2.

TKT silencing reversed epithelial-mesenchymal transition (EMT). (A) TKT regulated EMT molecules as determined by western blot assay. After transfection with TKT-shRNA or the vector plasmids, the cellular protein was extracted and subjected to western blot analysis. The expressions of EMT-associated molecules, including fibronectin, N-cadherin, IQGAP1, and r-catenin were evaluated. Actin protein levels were determined as an internal control for protein expression. (B) TKT regulated EMT molecules as examined by confocal microscopy. After transfection with TKT-shRNA or the vector plasmids, the cells were fixed and subjected to immunostaining analysis. These staining included antibodies against fibronectin with Cy3-conjugated secondary antibody (red imaging), N-cadherin with Cy3-conjugated secondary antibody (red imaging), or r-catenin with FITC-conjugated secondary antibody (green imaging). Cells were further stained with DAPI (blue imaging) to show the localization of nucleus. Cells were then observed by confocal microscopy.