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. 2014 Jul;20(7):1078–1089. doi: 10.1261/rna.045187.114

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© 2014 Jafarifar 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. — Binding of the PRPF31 protein to 5′ stem–loop RNA-15.5K protein complexes. (A) Labeled wild type (Wt) or mutant U4atac 5′ stem–loop RNAs were incubated with 15.5K protein and different amounts of PRPF31 protein. In each panel, lane 1 is RNA only; lane 2 is RNA plus 15.5K protein; lane 6 is RNA plus PRPF31 protein, and lanes 3–5 are reactions containing the listed fold excess of PRPF31 protein to the RNA-15.5K complex. The positions of free RNA, RNA-15.5K complex, and PRPF31-15.5K-RNA complex (dotted regions), are indicated in the figure. (B) Plot showing the percentage amounts of supershifted RNA-15.5K complex. Using ImageJ, the amounts of RNA shifted by 15.5K protein and the amounts of RNA supershifted (dotted region) by 15.5K + PRPF31 proteins were measured for each titration. The percentage of supershifted RNAs was calculated for each titration and normalized to wild type, which was set to 100%.