Fig. 3

Co-localization of the eIF4E isoforms with PBs and SGs during heat shock. The eIF4E1, 2, 3 proteins (green) were ectopically produced in fusion with GFP in U2OS cells. Nineteen hours after transfection, the cells were exposed to 41.7 °C for 30 min, fixed and assessed for eIF3B-stained SGs (red) and DDX6-stained PBs (blue). Co-localization of the particular eIF4E with SGs and 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 SGs and PBs. The eIF4E3_A (f) was recruited only to SGs, and eIF4E3_B (g) co-localized with neither SGs nor PBs. Approximately 50 cells transfected with either vector were observed in two independent biological replicates. Scale bar, 20 µm