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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 4. — RBPMS and RBPMS2 proteins bind (AC)₉ and (CAC)₆ repeats in electrophoretic mobility shift assays (EMSA). (A) Synthetic RNAs representing 18-nt di- or trinucleotide repeats were radiolabeled (10 nM), incubated with 0–10 μM RBPMS recombinant protein (FLAG-HA-His₆), and separated on 1% agarose gel. Binding conditions specified in Materials and Methods. The electrophoretic mobility shift observed for the (AAU)₆ oligoribonucleotide (*) may not be due to RBPMS binding (see Results). We did not observe binding for (AG)₉, (GGC)₆, (GCC)₆, A₃G₁₅, A₁₈, C₁₈, U₁₈ with the same conditions (data not shown). (B) Binding specificity for recombinant RBPMS2 protein. Conditions same as in A. (C) Definition of minimal number of (AC)_n repeats required for RBPMS binding. Conditions same as in A. (D) Coomassie-stained 15% SDS-PAGE indicates purity of recombinant RBPMS and RBPMS2 proteins.