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. 2010 Jan 13;84(7):3624–3630. doi: 10.1128/JVI.02362-09

FIG. 7.

FIG. 7.

RNase III suppresses RNAi-mediated gene silencing. (A) qRT-PCR analysis of GFP expression in Sf9 cells. The cells were mock transfected (mock), transfected with pIZ/GFP (GFP), transfected with pIZ/GFP plus GFP-specific dsRNA (GFP+RNAiGFP), transfected with pIZ/GFP plus GFP-specific dsRNA and infected with HvAV-3e (GFP+RNAiGFP+AV), transfected with pIZ/GFP plus GFP-specific dsRNA and infected with RNase III recombinant baculovirus (GFP+RNAiGFP+RNaseIII), or transfected with pIZ/GFP plus GFP-specific dsRNA infected with HvAV-3e DNA polymerase (orf1) recombinant baculovirus (GFP+RNAi GFP+Pol [control]). The mRNA levels were determined and compared by setting the level observed in silenced cells to 1.0. Error bars represent the standard deviations of averages from three replicates. (B) siRNA duplex degradation by RNase III. Two different siRNA duplexes were incubated in the presence of a control protein (Vn50; lanes 1 and 2) and RNase III (lanes 3 and 4). Reactions were run on a 15% urea-PAGE gel and stained with ethidium bromide.