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. Author manuscript; available in PMC: 2014 Apr 1.
Published in final edited form as: Nat Struct Mol Biol. 2013 Sep 22;20(10):1214–1220. doi: 10.1038/nsmb.2664

Figure 4. An mRNA that duplexes with another mRNA by 3' UTR base-pairing is targeted for SMD provided it is translationally active.

Figure 4

a, Diagram of IRE-Gl mRNA base-paired with FLUC-AS 3' UTR mRNA, which contains 3' UTR sequences engineered to be complementary to 3' UTR sequences of Gl mRNA. b, Western blot (WB) of lysates of HeLa cells stably expressing IRE-Gl mRNA and transiently expressing FLUC-AS 3' UTR mRNA (+) or, as a negative control, FLUC mRNA (−), the specified siRNA, and the reference MUP mRNA. Cells were cultured in the presence of either hemin or deferoxamine mesylate (Df) for 18-h prior to harvesting. c, Histogram representations of RT-sqPCR analyses of samples analyzed in b. The level of FLUC-AS 3' UTR mRNA was normalized to the level of MUP mRNA (Supplementary Fig. 4b), and the normalized level in the presence of Df and Control siRNA is defined as 100. d, as in c, but the level of IRE-Gl mRNA was normalized to the level of cellular SMG7 mRNA (Supplementary Fig. 4b). Error bars, s.e.m.; # of independently performed experiments = 3; *, P <0.05 (one-tail t-test).