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. 2011 Nov 1;204(Suppl 3):S911–S918. doi: 10.1093/infdis/jir343

Figure 1.

Figure 1.

The interferon inhibitory domain (IID) of EBOV VP35 binds to the endogenous double stranded RNA binding protein 76 (DRBP76) in the presence of dsRNA. A, Experimental outline of MBP-VP35-IID pull down. B, Recombinant MBP or MBP fused to wild-type or mutant VP35 IID was incubated overnight with NP40 buffer or 293T cell lysate in the presence (+) or absence (−) of pIC. Unique bands (labeled 1, 2, and 3) were excised and analyzed by mass spectrometry. Results from the protein database search indicated that protein bands 1, 2, and 3 corresponded to RNA Helicase A, nucleolin, and DRBP76 (also known at NFAR-1 and NF90), respectively. C, Primary sequence of the DRBP76 isoform described previously [26]. The underlined regions indicate the enzymatically digested peptides identified by tandem mass spectrometry. The bold-italic region represents the peptide sequence used to generate a DRBP76 specific antibody used in (D) that is common to 2 of the 6 isoforms. D, Western blot analysis from the same material used in (B) probed with an anti-DRBP76 antibody.