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. 2011 Mar;17(3):439–452. doi: 10.1261/rna.2475211

FIGURE 4.

FIGURE 4.

The effects of NSC218351 and NSC92218 in the dual luciferase assay. (A,C) BY4741 expressing Fluc with either an AUG or UUG start codon was treated with various concentrations of NSC218351 and NSC92218, respectively, and normalized to the internal Rluc control with an AUG start codon. The Fluc/Rluc ratio from each sample was then normalized to the appropriate DMSO-only control, so that each ratio equals 1 in the absence of compound (normalized FlucUUG, red circles; normalized FlucAUG, blue squares). Points are the averages of at least seven independent experiments ±SE. (B,D) Raw luciferase activity values with increasing concentrations of NSC218351 and NSC92218. Red symbols indicate luciferase values from pFuugRaug plasmid, and blue symbols indicate luciferase values from pFaugRaug counter-screening plasmid (Fluc, squares; Rluc, circles). Luciferase activites are normalized to the DMSO control values in each experiment, and the normalized values of at least six independent experiments are averaged (± average deviation). (E) Relative levels of Fluc/Rluc mRNA measured using RT-q–PCR, from strain BY4741 expressing FlucUUG and RlucAUG treated with 2 μM NSC92218 or 50 μM NSC218351. The Rluc mRNA levels were used to normalize the Fluc mRNA levels, and the Fluc/Rluc ratio of the DMSO sample was used to normalize the samples treated with compounds. Data are the averages of duplicate samples.