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. 2010 Nov 12;11(12):936–942. doi: 10.1038/embor.2010.164

Figure 2.

Figure 2

Role of the cap subunits in RNA recruitment. (A) Sodium dodecyl sulphate gel showing the reconstituted Exo9 (lane 1), Exo8 (lanes 2–4) and Exo7 (lanes 5–7) after size-exclusion chromatography. (B) Purified Exo9 (lane 1) and exosome complexes containing any two of the three cap subunits (Exo ΔCsl4, Exo ΔRrp4 or Exo ΔRrp40; lanes 2–4) form stable complexes that bind to Rrp44 (D551N RNase inactive mutant) in pull-down experiments (lanes 5–8). (C) RNA protection assay. The purified exosome complexes were incubated with a 60-nucleotide-long RNA molecule and unprotected RNA was degraded with RNase A/T1. RNA protection by exosomes without Rrp44 is shown in lanes 1–7 and the protection by Rrp44(D551N)-containing exosomes in lanes 8–14. WT, wild type.