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. 2016 Apr 1;12(4):e1005521. doi: 10.1371/journal.ppat.1005521

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© 2016 Nair et al

This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.

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Fig 7 — (A) Left panel: SDS-PAGE and Silver staining of Flag antibody immunoprecipitated samples from Huh7 cells transiently transfected with pCDNA5 Helicase-flag vector. Right panel: Western of pCDNA5 and pCDNA5 Helicase-flag vectors using anti-Flag antibody. (B) Left panel: SDS PAGE and Coomassie blue staining of indicated amount of purified bacterial GST-ORF4 protein. “*” indicates copurified unrelated protein band. Right panel: Western of 100ng purified GST-ORF4 protein using anti-ORF4 antibody. (C) Assay of RNA strand displacement by Helicase in the presence of GST-ORF4. “#”: heat denatured dsRNA. (D) Left panel: SDS-PAGE and Silver staining of Flag antibody immunoprecipitated samples from Huh7 cells transiently transfected with pUNO ORF2-flag vector. Right panel: Western of pUNO and pUNO ORF2-flag vector using anti-Flag antibody. (E) Left panel: SDS-PAGE and Silver staining of Flag antibody immunoprecipitated samples from Huh7 cells transiently transfected with pCDNA5 ORF4-flag vector. Right panel: Western of pCDNA5 and pCDNA5 ORF4-flag vector using anti-Flag antibody. (F) RNA strand displacement assay demonstrating the effect of Huh7 cell expressed ORF4 on HEV helicase enzymatic activity. Schematic at the right represents the structure of dsRNA and ssRNA.