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| Figure 2: Transfection of macrophages with the eIF2a-S51A mutant attenuated the E‾/B48 lipoprotein-induced inhibitory effect on global mRNA translational efficiency. Raw 264.7 cells were stably transfected with a plasmid vector expressing eIF2a-S51A (S51A) or an empty pEGFP vector. The pEGFP- (A and B) and S51A-transfected (C and D) cells were incubated with 20 µg/ml of E‾/B48 lipoproteins (B and D) or culture medium alone (A and C). Polysomal and nonpolysomal RNAs were separated on sucrose gradients. The optical densities at 254 nm were plotted as a function of gradient depth. Translation efficiency was expressed as the percentage of the integrated area under the polysome fraction of the curve relative to the total area under both the polysome and non-polysome fractions (E). |
