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. 2013 Aug 31;41(22):10086–10109. doi: 10.1093/nar/gkt777

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© The Author(s) 2013. Published by Oxford University Press.

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 9. — COX-2 gene activation does not require cleavage of the promoter transcript. (A) qPCR showing effect of RNA12 or RNA duplex containing scrambled sequence (12scr) on promoter RNA (−81/+56) levels. n = 3. (B) In vitro analysis of target cleavage by RNA12 and RNA12nc (12nc). Bases within RNA12nc that are mismatched relative to the promoter RNA are underlined. (C) Effect of RNA12nc and RNA12scr on COX-2 promoter RNA levels (−81/+56). RNAs were transfected at 25 nM. n = 3. (D) qPCR and western showing effect of RNA12, RNA12nc and RNA12scr duplex on COX-2 mRNA levels (upper, n = 2) and protein levels (lower). RNAs were transfected at 25 nM. (E) RIP showing recruitment of GW182 to COX-2 RNA (−81/+56) by RNA12nc (25 nM). Scheme showing recruitment of AGO2 and GW182 proteins after addition of RNA12nc to an intact promoter RNA. Error bars are SD. ***P < 0.001 (t-test) relative to MM.