Figure 2

Forced expression of TFF3 in MCF7 cells modulates the expression of epithelial, mesenchymal and metastatic-related gene markers. (A) qPCR analyses of MCF7 cells with either forced or depleted expression of TFF3 for mRNA levels of key genes functionally involved in migration, invasion, metastasis, and epithelial-mesenchymal transition (EMT). Change in gene expression is expressed as fold difference, respectively. Fold change values are representative of three independent biological experiments. (B) Western blot analysis was used to assess the protein levels of epithelial and mesenchymal markers in MCF7 cells with either forced or depleted expression of TFF3 as described in Methods. (C) Confocal microscopic visualization of TFF3, CDH1, and VIM expression in MCF7 cells with either forced or depleted expression of TFF3. The green colour indicates TFF3 expression, white colour indicates CDH1 expression and red colour indicates the positive VIM expression; and the blue colour indicates nuclei. Images were captured under oil immersion X600 magnification. Statistical significance was assessed by using an unpaired two-tailed Student's t test (P <0.05 was considered as significant) using GraphPad Prism 5. Columns are the mean of triplicate experiments; bars, ± SD.