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. 2016 May;22(5):773–781. doi: 10.1261/rna.055699.115

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© 2016 Grudzien-Nogalska 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 1. — Nudt3 down-regulation increases expression of mRNAs implicated in cell motility. (A) RNA levels of nine mRNAs implicated in cell migration tested in MCF-7 cells expressing nontargeting shRNA (Con^KD, open bar), shRNA against Nudt3 (Nudt3^KD, filled bar), or shRNA against Nudt4 (Nudt4^KD, gray bar) are shown. (B) Western blot analysis of Nudt3 protein levels in Con^KD or Nudt3^KD MCF-7 cells ectopically expressing GFP, Flag-tagged wild-type Nudt3 (Nudt3), or mutant Nudt3 (Nudt3^EE/QQ). The band for Flag-Nudt3^EE/QQ runs slightly faster than that for Flag-Nudt3 likely due to differences in protein charge by the introduction of glutamic acid residues into the mutant protein. (C) A subset of the mRNAs implicated in cellular movement derived from the 144 mRNAs detected to increase greater than or equal to twofold by RNA-seq analysis in Nudt3^KD are shown. (D) Transcript levels of the indicated mRNAs were determined by qRT-PCR in Con^KD or Nudt3^KD cells complemented with the indicated proteins. RNA levels in A and D are derived from at least three independent experiments and normalized to 18S rRNA and presented relative to the value in control cells set to one. Error bars represent ±SD. P-values are denoted by asterisks. (*) P < 0.05, (**) P < 0.01, (***) P < 0.001 (Student's t-test).