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. 2016 Jan;22(1):61–74. doi: 10.1261/rna.053447.115

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© 2015 Kini 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 6. — PABPC1 binds in the 5′ UTR of transcripts and negatively regulates translation. (A,C,E) Screenshots from the UCSC genome browser of PABPC1 CLIP tags in the 5′ UTR for Safb (A), Amd1 (D), and Ccnd2 (G) mRNA. (B,D,F) The native Safb (B), Amd1 (D), and Ccnd2 (F) 5′ UTRs (top) or mutant derivatives lacking the A-rich PABPC1 binding site cluster (bottom) were separately inserted in-frame with the firefly luciferase ORF in a standard expression vector. The PABPC1 binding site, corresponding to the CLIP tag cluster in (A, C, and E), is represented by the black rectangle within the 5′ UTR. (G) Quantification of firefly luciferase mRNA levels (qRT-PCR; light gray) and luciferase enzymatic activity levels (luciferase assay; dark gray) for the Safb (B), Amd1 (D), and Ccnd2 (F) 5′ UTR constructs. (*) P < 0.05; (**) P < 0.01, two-tailed t-test.