Figure 7. Disruption of cellular polarization and adhesion is associated with upregulation of p27^Kip1 level.

(A) Ventral prostates of both wild type (WT) and AKT1-Tg mice stained with Z0-1 antibody and imaged by confocal microscope. Scale bar, 50μM..

(B) Rat embryonic fibroblast cells stably transfected with either vector or Myr-AKT1 were cultured under adherent and suspension condition. Protein lysates were prepared and immunoblotted with antibodies directed against p27^Kip1 (upper panel), phospho-Akt (S473), phosho-GSK3 (α and β) (middle panel) and Tubulin (lower panel).

(C) Protein lysates were made from primary human epithelial cells (LEAR and LEKAR) cultured under suspension (Susp) or adherent (Ad) conditions. Immunoblot analysis using antibodies directed against p27^Kip1(upper panel), phospho-Akt (S473) (middle panel), and Tubulin (lower panel)

(D) Primary human epithelial cells (LEAR) were transduced with three different shRNAs against E-Cadherin and shGFP as control. Cell were harvested 2 days after post selection and total protein lysates were prepared. E-Cadherin (upper panel), p27^Kip1 (middle panel) and Actin (lower panel) western blot analysis was performed

(E) LEAR cells with E-Cadherin ShRNAs and control (ShGFP) were grown in plate and pictures were taken 2 days after post selection. Scale bar, 100 μM.