FIG 9.

trans-Complementation of defective virion assembly by cotransfection with replicon RNA. Huh7.5 cells were coelectroporated with either SGR-wt or SGR(Δ3D5A) in combination with one of the three intragenomic complementation replicons, Δ/wt, wt/Δ, or Δ/Δ. Cotransfection of cells with SGR(ΔGDD) and wt/wt(ΔGDD) was included as a negative control for both replication and transmission of replicon RNAs to recipient cells. (Left) RNA replication was assessed by measuring firefly and Renilla luciferase activities over 72 h; data were normalized to those at the 4-h time point (mean ± SD; n = 2). (Right) Transmission of infectivity was assessed by transducing all experimental groups with FBMJFH1(C-NS2) and performing a trans-encapsidation assay. Data represent luciferase values from recipient cells 48 h after inoculation with donor cell supernatants (mean ± SD; n = 2). For the purpose of comparison, values showing the percent transmission compared to that observed for the SGR-wt plus Δ/wt group are included.