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. 2012 Jul 23;7(7):e41355. doi: 10.1371/journal.pone.0041355

Figure 1. TBX2 induces epithelial-mesenchymal transition (EMT) in breast epithelial cells.

Figure 1

(A) Bright field images (40x magnification) of murine HC11 mammary epithelial cells stably transfected with pCDNA3 (+vector) or pCDNA3-TBX2 (+TBX2) expression plasmids showing morphological changes in TBX2-expressing HC11. (B) Western blot analysis using whole cell lysates from HC11+vector and HC11+TBX2 cells shows TBX2-induced downregulation of epithelial (Epi) and upregulation of mesenchymal (Mes) marker proteins. E-cad  =  E-cadherin; ß-cat  =  ß-catenin; N-cad  =  N-cadherin; Vim  =  Vimentin. Actin (ß-actin) was used as loading control. Fold changes in protein levels quantified by densitometry and normalized to Actin values are shown. (C) Bright field images (40X magnification) of human MCF10A mammary epithelial cells stably expressing pCDNA3 or pCDNA3-TBX2 reveal mesenchymal transformation of MCF10A+TBX2 cells. (D) Western Blot analysis shows that ectopic expression of TBX2 in MCF10A cells prompts a switch of EMT marker expression. (E) Immunofluorescence analysis of TBX2 (red) and EMT marker expression (40X magnification) shows loss of membrane-associated expression of epithelial (green) with a reciprocal gain of mesenchymal (red) marker expression in HC11+TBX2 cells as compared to HC11+vector control cells. Nuclei were stained with Hoechst 33258 (blue). (F) qPCR analysis comparing Tbx2 and EMT marker expression in HC11+vector and HC11+TBX2 cells. E-cad  =  E-cadherin; ß-cat  =  ß-catenin; ZO1 =  zona occludens 1; Dsp  =  Desmoplakin; N-cad  =  N-cadherin; Mmp3 =  matrix metalloprotease 3. Values were normalized to Gapdh. Fold changes as compared to vector control cells are shown. Error bars represent the mean ± SEM (n = 3; Student t-test). P values are indicated.