Skip to main content
. 2009 Oct 18;16(11):1134–1140. doi: 10.1038/nsmb.1680

Figure 6. Nsp1 induces cleavage at the IRES in Ren-EMCV-FF RNA.

Figure 6

(a) Schematic diagram of Ren-EMCV-FF RNA and control RNAs RNA1 and RNA2. (b,c) After incubation of RRL with GST, nsp1 (wt), or nsp1-mt (mt) for 10 min at 4 °C, Ren-HCV-FF RNA (b) or Ren-CrPV-FF RNA (c) was added to each sample and incubated at 30 °C for 25 min. Then RNAs were extracted and analyzed by northern blot using a digoxigenin-labeled probe that binds to the fLuc ORF. RNA lane in b, the untreated Ren-HCV-FF RNA; fLuc lane in c, the RNA encoding fLuc ORF; RNA, the untreated Ren-CrPV-FF RNA. (d) RRL was incubated with GST, nsp1 (wt), or nsp1-mt (mt) for 10 min at 4 °C. Ren-EMCV-FF RNA was added to each sample and incubated at 30 °C for 10 min. RNAs were extracted and analyzed by northern blot using a probe that binds to the rLuc ORF (rLuc probe) or fLuc ORF (fLuc probe). Samples in the first three lanes, including untreated Ren-EMCV-FF (RNA), RNA1 and RNA2, served as size markers.