Fig. 4

Co-localization of the eIF4E isoforms with PBs in control stress-free cells. The eIF4E1, 2, 3 proteins (green) were ectopically produced in fusion with GFP in U2OS cells. Nineteen hours after transfection, the cells were fixed and assessed for eIF3B-stained SGs (red) and DDX6-stained PBs (blue). No development of stress granules was observed. Co-localization of the particular eIF4E with PBs is demonstrated in the boxed area replicated in higher magnification on the right side of each panel and by the intensity profile measured along the dashed white line within the boxed area. Both eIF4E1 (a, b) and all three eIF4E2 (c–e) variants co-localized with PBs. No co-localization with PBs was detected for eIF4E3_A (f) or eIF4E3_B (g). Approximately 50 cells transfected with each plasmid were investigated in two independent biological replicates. Scale bar, 20 µm