Akt phosphorylation of T308 and S473 are independent events. Panel A.

<p>A bar diagram showing independence of phosphorylation of Akt at T308 (▪) and S473 (□) in kinase assays of the 1-LN cancer cells stimulated with α₂M* (50 pM/25 min) or insulin (200 nM/20 min). The bars are: (1) lipofectamine + buffer; (2) lipofectamine + α₂M* (50 pM/25 min); (3) Raptor dsRNA (100 nM/48 h) then α₂M*; (4) Rictor dsRNA (100 nM/48 h) then α₂M*; (5) lipofectamine + insulin; (6) Raptor dsRNA (100 nM/48 h), then insulin (200 nM/20 min); (7) Rictor dsRNA (100 nM/48 h) + insulin; and (8) scrambled dsRNA (100 nM/48 h) then insulin. Values are mean ± SE from three experiments and are expressed as fmol [³³P]-γ-ATP incorporated/mg cell protein. <b>Panel B.</b> Levels of p-Akt^T308 and p-Akt^S743 in cell lysates of 1-LN cells treated with α₂M* (50 pM/25 min) or insulin (200 nM/15 min). Representative immunoblots of p-Akt^T308 and p-Akt^S473 from three experiments of cells transfected with Raptor dsRNA or Rictor dsRNA as above are being shown.