Fig. 7

Akt isoforms differentially affect E-cadherin expression and localization. Cellular fractionation experiments were performed to characterize the subcellular distribution of E-cadherin in PANC-1 cells expressing (A) EGFP-K-Ras(V12), (B) EGFP, and (C) in H23 cells with Akt isoform depletion. Triton X-100 fractionation was performed to analyze the association of E-cadherin with the cytoskeleton [Triton insoluble] or as soluble protein [Triton soluble] and crude membrane preparations [P100] for localization within membranes. Total cell lysates were used to detect the total amount of E-cadherin [Total]. Protein lysates (30–50 μg) were analyzed by SDS-PAGE and Western blot procedure using the indicated antibodies. The bar graphs on the right show the relative E-cadherin protein amount estimated by densitometric quantification in each Akt-kd cell clone, in relation to an appropriate control protein, normalized to the corresponding cell clone transduced with a scrambled shRNA [Scr] set to 1. Mean ± SD (n = 3–8, one sample t-test, **** p ≤ 0.0001; *** p ≤ 0.001; ** p ≤ 0.01; * p ≤ 0.05) is shown