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. 2014 Jul;20(7):1014–1022. doi: 10.1261/rna.042648.113

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© 2014 Gudavicius 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 2. — FKBP25 interacts with nucleolin RRMs 1 and 2 in the presence of rRNA. (A) GST pulldown assays of GST, GST-FKBP25 N-terminal domain (NTD), GST-FKBP25 PPI and GST-FKBP25 FL against 6-His nucleolin RRM1-4 + RGG. Pulldowns were performed with and without rRNA. (B) GST pulldown assays using GST-NCL RRM1-2 (aa 290–480) (see Supplemental Fig. S4) against full-length 6His-FKBP25 in the presence and absence of RNase and supplemented with total RNA from HEK293 cells. (C) FLAG Immunoprecipitations from HEK293 cellular extract with and without pretreatment of RNase A. (D) Trizol extracted RNA from FLAG immunoprecipitations from HEK293 nuclear material electrophoresed on a denaturing formaldehyde-agarose gel. (E) Schematic of rDNA transcript. The position of primer sets used for qPCR is indicated below the schematic. (F) qPCR analysis of reverse transcribed RNA extracted from FLAG immunoprecipitations. GAPDH is used as an mRNA control.