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. 2005 Sep 12;170(6):913–924. doi: 10.1083/jcb.200504039

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Copyright © 2005, The Rockefeller University Press

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Figure 4. — eIF4E requires interaction with 4E-T for localization to P-bodies, whereas the other components of the eIF4F complex are excluded from P-bodies. (A) HeLa cells were transfected with HA-eIF4E wt, HA-eIF4E W73A, or HA-4E-T Y30A. Localization of proteins was studied by indirect immunofluorescence with anti-HA (Covance) and Alexa Fluor 594 anti–mouse IgG. (B and C) HeLa cells were transfected with GFP-Dcp1b and staining of endogenous eIF4GI (B) or eIF4A (C) was detected with anti-eIF4GI and Texas Red conjugated anti–rabbit IgG or with anti-eIF4A and Alexa Fluor 594 anti–mouse IgG. (D) HeLa extract (50 and 100 μg) was resolved by SDS-8% PAGE and a Western blot was performed with rabbit anti–eIF4GI polyclonal antibody. For eIF4A detection, proteins were resolved by SDS-10% PAGE and Western blot was performed with a mouse anti-eIF4A monoclonal antibody.