Mutation of phosphorylation sites in the PTEN C-terminus inhibits TGFβ-induced smad-independent pathways, but not the smad-dependent pathway in H358 cells.

Cell extracts from H358ON cells expressing Dox-dependent GFP, GFP-PTENWt, or GFP-PTEN4A in the absence or presence of Dox were harvested for analysis of the levels of total and phosphorylated for smad2 (A), Akt473 (B), Akt308 (B), and FAK (C) at the indicated periods after treatment with vehicle or TGFβ (1hour for smad2, 1hour for Akt473, 1hour for Akt308, and 24hours for FAK, respectively). A representative blot from three independent experiments is shown (top in A, B and C). The ratio of phosphorylated protein to total protein is presented as the intensity level relative to that in H358ON cells expressing Dox-dependent GFP treated with vehicle in the absence of Dox (bottom in A, B and C). Data shown represent the means ± SE. The experiment was repeated three times with similar results. *: p<0.05 N.S. indicates “not significant”.