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. 2014 May 24;16(4):365–374.e2. doi: 10.1016/j.neo.2014.04.004

Figure 5.

Figure 5

NHERF1 depletion induces displacement of apical PTEN and nuclear translocation of β-catenin in 3D spheroids. (A) Western blot of proteins (30 μg) from whole cell lysates with either Myc or PTEN A2B1 antibodies shows expression of Myc-tagged PTEN and PTEN-ΔPDZ mutant in Caco-2 cells. Endogenous PTEN is indicated by arrowhead. (B) IF of 3D structures developing from control and NHERF1 shRNA-depleted (shNHERF1) cells expressing Myc-tagged PTEN is shown. (C) IF analysis of 3D glands expressing Myc-tagged PTEN and PTEN-ΔPDZ shows apical PM localization of wild-type PTEN (arrowhead) as in B but not of PTEN-ΔPDZ that is partially localized to the nucleus (arrows). NHERF1 depletion delocalizes PTEN to the cytoplasm (right panels). (D) Confocal IF shows strong nuclear β-catenin (arrows) in some cells of NHERF1-depleted spheroids.