Figure 1

Zeb1 expression induces EMT/miR-200 repression and FAK pathway activation in previously non-invasive cells.

(a) Zeb1 expression causes a morphology change (middle panel) and is reversed upon re-expression of miR-200 (lower panel). Cells were stained with Integrin α6 (red), ZO-1 (green) and DAPI. (b) 393P cells with constitutive Zeb1 expression and 344SQ cells with constitutive miR-200 family expression grown on agarose and stained with phalloidin (red), DAPI and cortactin (green, lower panels). (c) Quantitative RT-PCR (left) and Western blot (right) analysis of EMT markers in the indicated cell lines. (d) Taqman RT-PCR for the miR-200 family members in the indicated cell lines. (e) In vitro Transwell migration and invasion assay for the 393P cell line panel. *p < 0.004, **p < 0.001 (f) Induction of miR-200a and b in H157 causes a morphology change. (g) Western Blot analysis of EMT markers in H157 + /− miR-200a and/or b. (h) In vitro Transwell migration and invasion assay for the inducible H157 cells. (i) Immunofluorescence of 393P_vec and 393P_ZEB1 cells stained for p-Src Y⁴¹⁸, p-FAK Y⁸⁶¹ and phalloidin (left); an average of 15–30 cells was counted for focal adhesions/cell with individual puncta considered a focal adhesion. Scale bar is 200 μm. (J) Western blot analysis of total cell lysates, cytosolic or membrane particulate fractions from 393P_vec and 393P_ZEB1 cells.