Figure 2.

Inactivation of TIF1γ expression enhances TGF-β1-induced EMT. MCF10A (A) and HMEC-TR (B) cells were knocked down using siRNA and treated or not with TGF-β1 for 96 h. To block any signalling arising from autocrine production of TGF-β, experiments without TGF-β were performed using the TβRI kinase inhibitor SB-431542. Phase-contrast images ( × 10) and immunofluorescence confocal acquisitions showing actin subcellular localization detected by phalloidin or indirect immunofluorescence using E- or N-cadherin-targeting antibodies. Scale bars, 26 μm. (C) Endogenous TIF1γ, Smad4, vimentin, N-cadherin and E-cadherin levels were determined by immunoblotting. GAPDH was used as a loading control. ctrl, control; EMT, epithelial-to-mesenchymal transition; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; siRNA, short-interfering RNA; S4, Smad4; TGF-β1, transforming growth factor-β1; TIF, transcription intermediary factor 1γ.