A. Membrane association of LASV Z wild-type and Z-G2A mutant. Huh7 cells were transfected with wild-type Z and Z-G2A mutant, respectively and 24 h post-transfection, flotation analysis was performed as described in Materials and Methods. Aliquots of the fractionated gradient were subjected to SDS-PAGE followed by immunoblotting using specific antisera against Z. Visualization was carried out with Odyssey™ Infrared Imaging System using a goat anti-rabbit secondary antibody labelled with Alexa680. B. VLP formation of Z-WT and Z-G2A. Cellular supernatants of Z-WT- or Z-G2A-transfected Huh7 cells were collected after 48 h, freed from cellular debris and then pelleted through a 20% sucrose cushion by ultracentrifugation (UC). UC-pellet containing VLPs and cell lysates as a control were dissolved in SDS-PAGE sample buffer and analyzed by SDS-PAGE and immunoblotting.