Fig. 3

Effect of PP1α and PP1γ silencing on AKT isoforms and AS160 in insulin sensitive and insulin resistant condition: Proliferated N2a cells were transfected with non-specific (scrambled) and PP1α and PP1γ specific siRNA. Post transfection cells were differentiated in the absence (MF; insulin sensitive) or chronic presence of 100 nM insulin (MFI; insulin resistant) for 3 days. Transfected N2a cells were treated with or without 100 nM insulin for 30 min, lysed and probed with relevant primary antibodies for immunoblotting. C and F Post insulin stimulation, lysates were subjected to immunoprecipitation using anti-Akt3 antibody. Bar represents relative change in A and D pAKT1 (Ser473) B and E pAKT2 (Ser474) C and F pAKT (Ser473) G and H pAS160 (Ser588) I and J pAS160 (Thr672). Experiments were executed three times and a representative western blot is shown. Data expressed are mean ± SE. ***p < 0.001 compared to Lane 1, ^###p < 0.001 compared to Lane 2, ^θθθp < 0.001 compared to lane 4 and ^δδδp < 0.001 compared to Lane 6. (A and D) AKT1, (B and E) AKT2, (C and F) AKT3, (G–J) AS160 was used as a loading control. Open bars: MF, filled bars: MFI, IP Immunoprecipitation IB Immunoblot, SC Scrambled