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. 2008 Feb 26;36(7):2434–2445. doi: 10.1093/nar/gkn093

Figure 6.

Figure 6.

Role of the uORF in the translational initiation control. Left, schematic representation of construct with uORF mutation. The internal promoter beginning constructs encoding each VEGF isoform or the VEGF minigene either with wild type uORF (pIP121mSPHA, pIP165mSPHA, pIP189mSPHA and pIPminiVEGF numbered 3), a mutated uORF AUG (pIP121mORFmSPHA, pIP165mORFmSPHA, pIP18mORFmSPHA and pIPminiVEGFmORF numbered 2) or a mutated uORF stop codon, which made the uORF in frame with the VEGF-HA (pIPmSOmSP121HA, pIPmSOmSP165HA, pIPmSOmSP189HA and pIPminiVEGFmSO numbered 1). Right, expression of AUG-initiated VEGF isoforms and uORF were analyzed by western immunoblotting using an anti-HA antibody and anti-β-actin to control loading of protein. RPA analyses were performed to normalize transfection with a vector specific probe to quantify mRNA from transfection and with a β-actin probe to control RNA quantitation.