Raptor immunodepletion and Akt^Thr308 phosphorylation.

<p><b>Panel A</b>. Effect of immunodepletion of Raptor in cell lysates of 1-LN cells stimulated with α₂M* (50 pM/20 min) or insulin (200 nM/20 min) on Akt^Thr308 phosphorylation. <b>Panel B</b>. Same as in <b>Panel A</b> except in cell lysates PDK1 was immunodepleted. Phosphorylation of Akt at Thr³⁰⁸ in <b>Panels A and B</b> was determined by kinase assay in Raptor immunoprecipitates from cells stimulated with: (1) buffer; (2) α₂M* or insulin; and (3) α₂M* or insulin + cell lysates of Raptor (<b>Panel A</b>) or PDK1 depleted cells (<b>Panel B</b>). Phosphorylation of Akt at Thr³⁰⁸ is expressed as fmol [³³P]-γ-ATP/mg cell protein and is the mean ± SE from three independent experiments.