Therapeutic suppression of translation initiation factor eIF4E expression reduces tumor growth without toxicity
J. Clin. Invest. Jeremy R. Graff, et al. 117:2638 doi:10.1172/JCI32044 [Go to this article.]

Figure 3
Reduction of eIF4E and global protein synthesis. (A) The human breast cancer cell line MDA-MB-231 was transfected for 72 hours with 4E-ASO4, 4E-ASO2, or the mismatch ASO control at the indicated concentrations. Cells were labeled 72 hours after transfection with ³⁵S-methionine/cysteine (Promix). As a positive control for blocking total protein synthesis, cells were pretreated with cycloheximide (CHX) 4 hours before the addition of Promix. Equal protein was loaded per lane on an SDS-PAGE gel, electrophoresed, dried, and exposed to a phosphorimager screen overnight. (B) RT-PCR analyses for eIF4E expression normalized to total RNA as determined by Ribogreen stain are depicted for MDA-MB-231. (C) ³⁵S incorporation per lane was normalized to total RNA from the same sample. Data represent 2 separate experiments in MDA-MB-231.