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(A) TR-FRET assays were developed for the quantification of either p-AKT(Ser473) or p-cSTKpep-3. (B) p-AKT(Ser473) and (C) p-cSTKpep phosphorylation was quantified after treatment with Staurosporine (open triangles), Wortmannin (open squares), LY294002 (open circles) or DMSO (closed squares). Experiments were performed in 384-well plates after transient co-transfection of AKT-His and cSTKpep-3 into HEK293 cells.
