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. 2010 Jul 7;38(20):7260–7272. doi: 10.1093/nar/gkq611

Figure 3.

Figure 3.

Differential effects of optimized upstream start codons on translation of downstream ORFs. (A) Diagram indicating the location of the optimized (opt) (ccAccAUGG) p10 or p17 start codons (top panel). RNA-transfected QM5 cells and cell lysates were harvested and assayed for luciferase activity (bottom panel) as described in Figure 2C. (B) Diagram depicting a capped RL gene (shaded rectangle) with a β-globin 5′-UTR, either with (β-ORF-RL) or without (β-RL) an upstream ORF (black rectangle) (top panel). RNA-transfected QM5 cells were harvested and analyzed for luciferase activity (bottom panel) as described in Figure 2C. (C) The σC-RL constructs represented in (A) were 5′-terminally extended through the addition of the β-globin 5′-UTR, and RNA-transfected cells were harvested and analyzed for luciferase activity (bottom panel) as described in Figure 2C. All results are reported as mean ± SE (n = 4–5) of the relative level of RL (B) or σC-RL (A and C) activity, normalized to the parental σC-RL (A), β-RL (B) or β-σCRL (C) constructs. Expression levels significantly different from the parental constructs (***P < 0.001 and **P < 0.01) are indicated.