Figure 7

Envelope glycoprotein incorporation efficiency and infectivity of henipavirus F and G bearing lentivirus particles. The panel of expression plasmids encoding the NiV and HeV F glycoprotein cytoplasmic tail truncation mutants and/or their G glycoprotein partner together with the HIV-1 backbone pNL4-3-Luc-E-R⁺ were transfected into 293T cells. The pseudovirus containing cell culture supernatants were collected 36 hr post-transfection, filtered with a 0.45 μm filter and purified through a 25% wt/vol sucrose cushion. The preparations of pseudovirions were normalized by assaying p24 content and then used to infect permissive 293T target cells. (A) Infection assay with the various NiV F cytoplasmic tail deletion mutants. (B) Infection assay with the various HeV F cytoplasmic tail deletion mutants. Error bars indicate the standard error of the mean from triplicate wells. (C) Incorporation of the various NiV and HeV F glycoproteins into the lentivirus-based pseudovirions. Equal amounts of particles, based on p24 content, were lysed and subjected to SDS-PAGE and Western blot analysis to assess the levels of incorporation of the F glycoproteins. Mock is processed supernatant prepared from cells not producing pseudovirions. This experiment was performed twice and representative experiment is shown in the figure.