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. 2014 Jul;20(7):1090–1102. doi: 10.1261/rna.045005.114

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© 2014 Farazi 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 5. — RBPMS binds PAR-CLIP targets in EMSAs. (A) 21-nt synthetic RNAs representing clusters from mRNA targets identified by PAR-CLIP (NDUFA6, UBE2V1, SRM, and ETF1) were radiolabeled (10 nM), incubated with 0–10 μM recombinant RBPMS or RBPMS2 (FLAG-HA-His₆), and separated on 1% agarose gel. The RNA sequences are shown on top, with nucleotides representing the RRE boxed. Residues in red illustrate point mutations of the synthetic RNAs. (B) Dissociation constant (K_d) determination for 21- and 15-nt synthetic RNAs representing the wild-type and mutant NDUFA6 cluster. (C) Binding study of RBPMS deletion proteins (His₆) to synthetic (AC)₉ RNA defines additional regions contributing to RNA binding. C-terminal deletion proteins are specified; N-terminal deletion protein (amino acids 21–196) required different binding conditions and could not be directly compared (data not shown).