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. 2019 Nov;25(11):1549–1560. doi: 10.1261/rna.071811.119

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© 2019 Musalgaonkar 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. — Truncation of L1 stalk RNA leads to a 60S export defect. (A) Cartoon of 25S rRNA for WT and L1 stalk truncation showing expected lack of Rpl1 binding when RNA was truncated. (B) Plasmids constructs expressing WT (pAJ1181) or L1 stalkΔ (pAJ3605) rRNA were transformed into AJY1185 (rDNAΔ, 35S URA3 2µ) and complementation was tested on 5-FOA media. (C) Fluorescence in situ hybridization and microscopy using oligo (AJO628) hybridizing to a unique tag in 25S rRNA expressed from plasmid constructs for WT (pAJ1181) and L1 stalkΔ (pAJ3605) in strain BY4741. (D) Sucrose gradient sedimentation and northern blot analysis using oligo (AJO628) against a unique tag on rRNA expressed from WT (pAJ1181) or L1 stalkΔ (pAJ3605) rRNA. Total 25S and 18S rRNAs were detected using oligos AJO192 and AJO190, respectively.