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. 2013 Jul 2;8(7):e66157. doi: 10.1371/journal.pone.0066157

Figure 3. CHRNA5 5 .

Figure 3

′UTR harbours a functional uORF; CHRNB3 uORFs are not involved in translational control. A: Luciferase assay of CHRNA5 5′UTR resulted in significant increase in protein expression when switching off the uORF. Fold change of firefly activity normalized to pGl4.10 +TK, is illustrated for the five different constructs; 1, pGl4.10+TK; 2, CHRNA5-1; 3, CHRNA5-2; 4, CHRNA5-3; 5, CHRNA5-4. B: The uORF start codon of CHRNA5 did not initiate translation as efficiently as the firefly start codon. Fold change of firefly activity normalized to pGl4.10 +TK, is illustrated for the five different constructs; 1, pGl4.10+TK; 2, pGL4.10; 3, CHRNA5-2; 4, CHRNA5-5; 5, CHRNA5-6. C: Relative qPCR for CHRNA5 5′UTR showed no significant differences of mRNA amount when comparing intact with deleted uORF. However, significant differences were assessed for the transcription of the two SNP allele variants. Fold change of firefly mRNA normalized to pGl4.10 +TK, is illustrated for the five different constructs; 1, pGl4.10+TK; 2, CHRNA5-1; 3, CHRNA5-2; 4, CHRNA5-3; 5, CHRNA5-4. D: Luciferase assay of CHRNB3 5′UTR showed no significant results. Fold change of firefly activity normalized to pGl4.10 +TK, is illustrated for the four different constructs; 1, pGl4.10+TK; 2, CHRNB3-1; 3, CHRNB3-2; 4, CHRNB3-3. Error bars represent ± SEM of 3 biological replicates. Asterisks indicate significant differences (p<0.05).