Fig. 1

Downregulation of Foxf2 attenuates TGFβ-induced EMT. a Phase-contrast micrographs of NMuMG cells stably expressing a control shRNA (shCtrl) or shRNAs against Foxf2 (shFoxf2 703, shFoxf2 704) treated with transforming growth factor (TGF)β for the times indicated. Scale bar = 100μm. b Foxf2 knockdown efficiency was determined by quantitative RT-PCR in NMuMG cells stably infected with shCtrl, shFoxf2 703, or shFoxf2 704 and treated with TGFβ for the times indicated. Values were normalized to RPL19 and presented as fold changes compared with untreated shCtrl NMuMG cells. c Loss of E-cadherin expression during TGFβ-induced EMT depends on Foxf2. E-cadherin mRNA levels in shFoxf2 and shCtrl-transfected NMuMG cells were determined by quantitative RT-PCR. Values were normalized to RPL19 and reported as fold changes compared with untreated shCtrl NMuMG cells. d Knocking down Foxf2 leads to a sustained expression of cell junction components. Immunoblotting analysis for the epithelial markers E-cadherin and ZO-1 as well as the mesenchymal markers Ncam1, N-cadherin, and fibronectin in shFoxf2 knockdown and shCtrl NMuMG cells treated with TGFβ for the times indicated. Actin was used as a loading control. Data are shown as mean ± SEM of three independent experiments. Statistical values were calculated using a paired two-tailed t test. *p ≤ 0.05; **p ≤ 0.01; ***p ≤ 0.001