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. Author manuscript; available in PMC: 2008 Jan 12.
Published in final edited form as: Cell. 2007 Jan 12;128(1):129–139. doi: 10.1016/j.cell.2006.11.039

Figure 5. NEDD4-1 potentiates cell transformation in a PTEN-dependent manner.

Figure 5

(A) NEDD4-1 regulates AKT phosphorylation. RNAi plasmid iLacZ or iN4A was transfected into 293 cells by electroporation. Immunoblotting against NEDD4-1, PTEN, phospho-AKT (p-Akt), total AKT, and β-tubulin was performed as indicated.

(B) Overexpression of NEDD4-1 in p53−/− primary MEFs decreased endogenous PTEN level. The MEFs were transfected with vector control or NEDD4-1 (N4-HA) as indicated by electroporation. The cells were lysed 24 hours later. N4-HA and endogenous PTEN were detected by immunoblotting. β-tubulin was blotted as the loading control.

(C) Overexpression of NEDD4-1 promotes Ras-induced cell transformation in p53−/− primary MEFs. The p53−/− primary MEFs were infected with retroviruses encoding for no ectopic protein (Vector), Ras, and/or NEDD4-1 (N4), as indicated. Soft-agar colony formation assays were performed as described in Experimental Procedures. The pictures showing the typical colony formation assay result.

(D) Overexpression of NEDD4-1 promotes cell transformation in a PTEN-dependent manner. The colony formation assays were performed in both p53−/− primary MEFs and p53+/−Pten−/− primary MEFs. The plot represents the results from three independent experiments with standard deviation.