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. 2010 Nov 24;5(11):e15057. doi: 10.1371/journal.pone.0015057

Figure 4. Effect of LNA-AUG1 is independent of different 5′ leader lengths.

Figure 4

A. CAT-FLAG protein expression from AUG1 normalized with that from AUG2 in (CAA)n CAT-FLAG mRNAs. COS-7 cells were transfected with plasmid constructs that express (CAA)4, (CAA)10, or (CAA)16 CAT-FLAG mRNAs, along with a control plasmid that expresses the control FLAG-Luc2 mRNA. LNA oligonucleotides (100 nM) were cotransfected into COS-7 cells as indicated. Expression levels of the two CAT-FLAG proteins were analyzed as in Figure 2. B. CAT-FLAG protein expression from AUG1 normalized with that from AUG2 in COS-7 cells transfected with a plasmid construct that expresses 0.25X, 0.5X, or 1.0X β-globin CAT-FLAG mRNA. At least three independent experiments were performed for final quantification of the immunoblot with error bars indicating standard deviations. For these experiments, cotransfection of cells with LNA-AUG1 resulted in significant differences in relative AUG codon usage (one-sided t-test: * p<0.05; ** p<0.01); no significant change in relative AUG codon usage was observed in cells cotransfected with LNA-C (p>0.05).