Figure 4

Role of Akt in PPP2CA overexpression-mediated suppression of β -catenin and NF- κ B. (A) PC3-PPP2CA cells were transiently transfected with constitutively active PKB mutant or control plasmid for 24 h, and LNCaP-shPPP2CA cells were pre-treated (for 1 h) with PI3K/Akt inhibitor (LY294002). Subsequently, cells were co-transfected with TOPflash/FOPflash/NF-κB luciferase promoter–reporter constructs along with control plasmids for 24 h and luciferase activities were measured. Bars represents mean±s.d. (n=3), *P<0.01. (B) Following transfection with active PKB mutant or PI3K inhibitor treatment, nuclear and total proteins were prepared and effect on localisation of β-catenin and NF-κB (in nuclear protein; upper panel) and on p-Gsk3-β/Gsk3-β and p-IκB-α/IκB-α (in total protein; lower panel) was examined by immunoblot analysis. Laminin (for nuclear protein) and β-actin (for total protein) were used as loading controls.