Essential role of host translation regulatory factors in the assembly of viral translation/replication complex. Essential role of host translation regulatory.

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Essential role of host translation regulatory factors in the assembly of viral translation/replication complex. Essential role of host translation regulatory factors in the assembly of viral translation/replication complex. (A) QRT-PCR of sense and antisense RNA of mock, WT HEV, and GAA HEV infection in Huh7 cells expressing various siRNAs/shRNA, as indicated. Relative RNA levels (normalized to that of GAPDH) are represented as means ± SEM. (B) QRT-PCR of sense and antisense RNA of g-1 HEV-infected ORF4-Huh7 cells expressing various siRNAs/shRNA, as indicated. Relative RNA levels (normalized to that of GAPDH) are represented as means ± SEM. (C) (Upper panel) HEV RdRp assay using the indicated siRNA/shRNAs and RdRp-Flag-transfected cells. (Lower panel) Western blot analysis of aliquots of RdRp protein used in the assay, performed using Flag antibody. (D) (Upper panel) HEV RdRp assay using affinity purified RdRp-Flag and the indicated viral proteins from Huh7 cells. (Lower panel) Western blot analysis of aliquots of RdRp protein used in the assay, performed using Flag antibody. (E) Myc-agarose pulldown assay of the indicated siRNAs/shRNA and RdRp-myc-transfected Huh7 cells, followed by Western blot analysis of various viral and host factors, performed using the indicated antibodies. Chandru Subramani et al. mSystems 2018; doi:10.1128/mSystems.00135-17