Fig. 8.

eIF4G interacts with eIF4E3_A but not with eIF4E3_B. a Coomassie blue stained gel demonstrating immunoprecipitation of the ectopically expressed GFP-eIF4E3_A from the HEK293 cell lysate using a GFP-Trap approach. M PageRuler™ Prestained Ladder (Thermo Scientific); INPUT whole cell lysate; eIF4E3_A-IP proteins co-immunoprecipitating with GFP-eIF4E3_A. b MS analysis of the proteins co-immunoprecipitating with eIF4E3_A. Gel slices are numbered as in panel A. c Western blots of proteins co-immunoprecipitating with eIF4E1_1, eIF4E3_A and eIF4E3_B transiently expressed in GFP fusion in HEK293 cells using GFP-Trap agarose beads (GFP-TRAP). Membranes were developed with anti-GFP (detecting eIF4E-GFP fusion proteins), anti-eIF4G and anti-β-actin antibodies. INPUT lines including β-actin and eIF4G served as a loading control. Lysate from non-transfected HEK293 cells was used as a negative control (mock). The absence of detectable β-actin on GFP-Trap beads shows no contamination of non-specifically bound proteins in the samples