Fig. 3
From: Monitoring flux in signalling pathways through measurements of 4EBP1-mediated eIF4F complex assembly
a Western blot analysis of endogenous phosphorylation status of AKT and S6 kinases in non-transfected HEK293 cells treated with either PP242, Torin, Rapamycin or the MNK1/2 inhibitor CGP57380. b Western blot analysis of endogenous 4EBP1 phosphorylation status in non-transfected HEK293 cells treated with either the dual mTORC1/2 active site inhibitors PP242 and Torin, or the allosteric inhibitor of mTORC1 Rapamycin c PANC1 cells, which contain low levels of endogenous 4EBP1 were co-transfected with the NanoBit eIF4E:eIF4G604–646 system with differing combinations of 4EBP1 mutant constructs with different sensitivities to mTORC1-mediated phosphorylation. Each transfection combination was normalised to its own DMSO treatment control. The molecular mass of the protein marker is indicated in kilodaltons