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. 2021 Mar;27(3):335–342. doi: 10.1261/rna.076885.120

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© 2021 Watson et al.; Published by Cold Spring Harbor Laboratory Press for the RNA Society

This article is distributed exclusively by the RNA Society for the first 12 months after the full-issue publication date (see http://rnajournal.cshlp.org/site/misc/terms.xhtml). After 12 months, it is available under a Creative Commons License (Attribution-NonCommercial 4.0 International), as described at http://creativecommons.org/licenses/by-nc/4.0/.

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FIGURE 3. — Roadblock-qPCR yields accurate measurements of mRNA half-life. (A) Reporter mRNA stability analysis using transcription shutoff with doxycycline. HEK-293T cells stably expressing a dox-off Renilla were treated with 1 µg/mL doxycycline for 0–8 h. Reporter mRNA levels were measured at the indicated times using qPCR. Error bars are SD, n = 3. (B) Reporter mRNA stability analysis using Roadblock-qPCR. RNA was isolated from the same HEK-293T cells expressing Renilla mRNA from A, now treated with 4sU for 0–8 h, then treated with NEM and reverse transcribed. Reporter mRNA levels were measured using qPCR. Error bars are SD, n = 3.