Skip to main content
. 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 3. mRNA–mRNA duplexes create SBSs and trigger SMD.

Figure 3

a, Western blot (upper) or RT-sqPCR(lower) of lysates of HeLa Tet-Off cells that had been transiently transfected with the specified siRNA (see Supplementary Fig. 3c for data using alternative siRNAs) in the presence of 2 μg/ml of doxycycline (DOX) and, after removing doxycycline 48-h later, with pTRE-FLUC-SOWAHC 3' UTR, which contains a doxycycline-repressible promoter, and the phCMV-MUP reference plasmid. The removal of DOX elicits a transcriptional burst in FLUC-SOWAHC 3' UTR mRNA production. A fraction of cells was harvested after an additional 4-h (time 0) and analyzed. b, Plot of RT-sqPCR data using samples shown in a that were further processed as follows. At time 0, 2 μg/ml of doxycycline were added to the remaining cells. Additional fractions of cells were harvested at the specified times thereafter. RT-sqPCR analyses were essentially as in Fig. 1b. As in Fig. 1d, the levels of exogenous FLUC-SOWAHC 3' UTR mRNAs were comparable to the level of cellular SOWAHC mRNA. Error bars, s.e.m.; # of independently performed experiments = 3; P < 0.05 (Chi-squared test).