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. 2010 Dec;12(12):1013–1022. doi: 10.1593/neo.10780

Figure 2.

Figure 2

NHERF1 depletion induces EMT-like changes and nuclear translocation of β-catenin in polarized Caco-2 cells. (A) Confocal IF analysis of a Caco-2 monolayer with NHERF1 (red) and E-cadherin (green) antibodies. XZ image (top) shows NHERF1 in the microvilli at the apical PM, and the XY middle-plane section (bottom) shows E-cadherin at the lateral cell-cell junctions. The arrow indicates the plane of XZ section. (B) Western blot of whole cell lysates showing NHERF1 depletion by two shRNAs (sh1 and sh4) and opposite variations of epithelial (E-cadherin) and mesenchymal (fibronectin) markers in NHERF1-depleted cells. Proteins were loaded at 30 µg per lane. (C) Crystal violet staining showing EMT-like morphologic changes in NHERF1-depleted cells compared with control (vector) cells. (D) Confocal IF with indicated antibodies shows nuclear accumulation and lateral PM effacement of β-catenin in NHERF1-depleted cells compared with vector control cells. The densitometric analysis of β-catenin intensity for control and NHERF1-depleted cells is shown. n = 25. *P < .05, ***P < .001 versus vector.