Fig. 2.
Activation of PI 3-kinase and Akt by NPM-ALK.
The 5 × 106 Ba/F3 cells, stably transfected with empty vector (∅) or NPM-ALK, were starved for 4 hours in medium containing 0.5% FCS without IL-3. (A) PI 3-kinase activity was measured by incubation of antiphosphotyrosine (4G10) immunoprecipitates with phosphatidylinositol and P32-γg-ATP. P32-labeled–enzymatic products of PI 3-kinase were resolved by thin-layer chromatography (TLC) using a silica gel plate as described in “Materials and methods,” and visualized by autoradiography. (B) Cell lysates were subjected to 7.5% SDS-PAGE and immunoblotted using a goat polyclonal anti-Akt antibody. Mobility shift of Akt indicates its activation.
