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Figure 3 | Molecular Cancer

Figure 3

From: Response to mTOR inhibition: activity of eIF4E predicts sensitivity in cell lines and acquired changes in eIF4E regulation in breast cancer

Figure 3

Translational efficiency specified by a structured 5'UTR reporter responds to changes in eIF4E activity. A) Reporters were constructed to express mRNAs containing the GFP reading frame preceded by a control 5'UTR lacking regulatory motifs (control) or a sequence predicted to form a stable stem-loop structure (structured). MCF7 cells were transiently transfected with equal copy numbers of either control or structured reporters along with either empty expression vector (-) or vector to allow over-expression of eIF4E (+). GFP protein and mRNA were quantified by flow-cytometry and real-time PCR respectively. Translational efficiency (protein synthesised per unit mRNA) is presented relative to the control. Data points represent means (+/-standard deviations) of values from three separate wells of cells within a representative experiment. B) Expression of exogenous proteins was confirmed within cells transfected with control GFP reporter and vector to allow over-expression of eIF4E as shown by Western blot analysis for the HA-epitope tag; exogenous GFP and eIF4E both include this epitope. Note that the small reduction in GFP protein associated with eIF4E-transfection does not result from a change in translational efficiency (see Panel A).

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