Fig. 5
From: Monitoring flux in signalling pathways through measurements of 4EBP1-mediated eIF4F complex assembly
HEK293 cells transfected with the NanoBit eIF4E:eIF4G604–646 system and treated with compound titrations of either a the AKT inhibitors, Ipasertib and AZD5363, b the ERK1/2 inhibitors, Ulixertinib and SCH772984 or c the mTORC/PI3K dual inhibitor BEZ235 for 4 h, and then measured for luciferase activity. d Table displaying the IC50 values determined for each compound titrated into the live cell NanoBit eIF4E:eIF4G604–646 PPI system. e HEK293 cells were co-transfected with NanoBit eIF4E:eIF4G604–646 system and either siRNA Ctrl or siRNA 4EBP1. Cells were then treated BEZ235 and luciferase activity assessed as in a, b and c. Four parameter models were fitted to the experimental data using non-linear regression to derive the IC50 values (Prism, GraphPad Ltd.). All values represent mean ± SD (n = 3). Data was normalised to DMSO controls