Fig. 5

Purified mTORC2 phosphorylates FLNA at Ser2152 in vitro. a In vitro kinase assay of mTORC2 purified from U87vIII cells with FLNA and AKT as substrates. Level of phosphorylated FLNA (Ser2152) and phosphorylated AKT (Ser473) were increased in the presence of mTORC2 and ATP. Negative control groups contain either no purified complex or no ATP. Activity of mTORC2 is maximal when Mn²⁺ is used instead of Mg²⁺. b In vitro kinase assay of mTORC2 purified from U87vIII cells with FLNA as a substrate. Two concentrations of PP242 (+ = 1.25 μM; ++ = 5.0 μM) and one concentration of IPA3 (+ = 40 μM) were added into three samples to inhibit the kinase activity of mTORC2. Levels of pFLNA and pAKT when treated with PP242 are decreased compared to ones without PP242