FIG 7.

trans-Complementation of NS5A virion assembly functions. (a) Expression of NS proteins in Huh7.5 cells at 24 h postransduction with baculoviruses encoding LacZ (lane 1), NS5A (lane 2), and NS3-5A (lane 3). (b) Transient replication of SGR, SGR(Δ3D5A), and SGR(ΔGDD) following transfection into Huh7.5 cells. Luciferase activity was measured at 4 and 72 h (mean ± SD; n = 2). (c) Transmission of replicons from donor to recipient Huh7.5 cells by trans-encapsidation assay. Cells were electroporated with the indicated SGR RNAs and then transduced after 24 h with baculovirus expressing the HCV core to NS2 region (C-NS2) alone or with baculoviruses expressing NS5A or NS3-5A for a further 6 h. At 48 h postransduction, supernatants were harvested and used to inoculate naive Huh7.5 cells. Luciferase activities were assayed at 48 h after inoculation of cells (mean ± SD; n = 2). Baculovirus expressing LacZ was used as a negative transduction control. For the purpose of comparison, values show the percent transmission compared to that observed for SGR-wt plus C-NS2.