Fig. 4
From: Lymphocytic choriomeningitis arenavirus utilises intercellular connections for cell to cell spread
Rescue of a recombinant LCMV variant with GP-1 FLAG tag alongside Z HA tag (rLCMV-GP1-FLAG-Z-HA). (A) Schematic of LCMV S and L segments with respective tag insertions. (B) BSR-T7 cells were transfected with cDNAs expressing a LCMV S segment encoding a FLAG tagged GP-1 (S-FLAG segment), an L segment encoding a HA tagged Z (L-HA segment), as well as NP and L open reading frames (support plasmids, L + and NP+) and a plasmid expressing T7 RNA polymerase. At 5 days post transfection (5 dpt), cellular supernatants were transferred to BHK cells, and at 3 days post infection (3 dpi), cell lysates were analysed by western blotting using NP antisera, with viral supernatants harvested and subject to focus forming assays (FFA). (C) Western blot analysis of transfected BSR-T7 and infected BHK-21 cell cultures confirmed rLCMV-GP1-FLAG-Z-HA rescue (alongside all other mutants), using antisera specific for LCMV NP and GAPDH as loading control.
