Fig. 5.

DAP5 and eIF3d are both required for translation of DAP5-dependent mRNAs. a Overexpression of 4E-BP1 by stable cDNA transformation in MDA-MB-231 cells silenced with siRNAs: non-silencing (NSi), DAP5, eIF4GI, eIF3d, DAP5 + eIF4GI, and DAP5 + eIF3d. b Cap chromatography of cell lysates with or without 4E-BP1 cDNA overexpression. Representative immunoblots show almost total sequestration of eIF4E by overexpressed 4E-BP1. c Representative immunoblots of MDA-MB-231 cells silenced with NSi or DAP5 siRNAs as in (a) probed for proteins whose mRNAs were found to be reduced in polyribosomes by DAP5 silencing (Table 1). Lamin A, AUF1, and GAPDH served as DAP5-independent controls. Results are representative of three independent experiments