Fig. 2.

Differential temporal- and dose-dependent increase in complemented FL activity by the PI-3K inhibitor LY294002 and Akt kinase inhibitor perifosine in 293T cells stably transfected with ASA (293T/ASA) or AST (293T/AST). a 293T/AST cells were treated with different concentrations of LY or carrier control for 6, 12, and 24 h prior to analysis of complemented FL activity by bioluminescence imaging of intact cells. Quantitation of bioluminescence signals in 293T/ASA and 293T/AST stable cells treated with LY was performed as follow: Total flux was determined by drawing ROI over each well with 5% border detection limit. Protein content in each well was determined upon cell lysis. Complemented FL activity in each well was normalized for protein content, averaged first and then normalized to carrier control treated cells. Each data point represents the average normalized complemented FL activity±SEM. *p<0.05 relative to ASA cells treated with same concentrations of LY for the same duration. b Increase complemented FL activities in 293T/ASA and 293T/AST cells upon treatment with Akt kinase inhibitor perifosine were determined as in a, except 0.1% ethanol was used as the carrier control. Quantitation of bioluminescence signals was performed as described (a). *p<0.05 relative to ASA cells treated with same concentrations of perifosine for the same duration.