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. 2017 Mar 21;12(3):e0173980. doi: 10.1371/journal.pone.0173980

Fig 2. Relationship between readthrough-mediated stabilization and downstream ORF length.

Fig 2

(a) NanoLuc or HIV RNase H sequences were added to dual-fluorescent reporter mRNAs in the indicated positions. (b) Decay assays of mRNAs in which UGA or UAA TCs following the GFP ORF were modulated by the MLVPK and NanoLuc (Nluc) or HIV RNase H (HIV RH) sequences were added as indicated. RNA half-lives corresponding to best-fit lines to semi-log plots of RNA abundance data and 95% confidence intervals are listed (n = 3). Readthrough efficiencies (mean +/- SD; n = 3) for each construct are indicated below the blot panels. (c) Semi-log plot of mRNA decay assays shown in B. Error bars indicate +/- SD (n = 3); p-values derived from ANCOVA analysis. (d) Decay assays of mCherry-NanoLuc reporter mRNAs as in A. Cells were treated with control siRNAs (siNS) or UPF1 siRNAs (siUPF1) as indicated. Half-lives are represented as in B. (e) Quantification of mRNA decay assays shown in D; note reduced duration of timecourse vs. C (360 min.). Error bars indicate +/- SD (n = 3); p-values derived from ANCOVA analysis.