Figure 4
From: Nck1 depletion induces activation of the PI3K/Akt pathway by attenuating PTP1B protein expression
Enhanced basal Akt phosphorylation in HepG2 cells depleted of Nck1 is dependent on PI3K, and correlates with increased total and p85-associated pY proteins. Overnight serum-starved HepG2 cells transfected with control or Nck1 siRNA were exposed to 10 μm of the PI3K inhibitor LY294002 for 30 min (A), untreated (B) or pretreated with 100 μm of pervanadate (PV) for 5 min (C) before harvesting. Equal amount of proteins from total cell lysates were subjected to immunoblot with indicated antibodies. Data are representative of three independent experiments performed in duplicate or triplicate and showing similar results. Total cell lysates prepared from HepG2 cells transiently transfected with control or Nck1 siRNA were subjected to anti-p85 (regulatory subunit of PI3K) immunoprecipitation followed by immunoblot using indicated antibodies (D). Data are representative of three independent experiments showing similar results.