Figure 1

Henipavirus F and G bearing pseudotyped lentivirus particles. (A) Infection assay with NiV F and G glycoprotein bearing virus particles. Virus particles were prepared in 293T cells by co-transfecting the pNL4-3-Luc-E-R⁺ HIV-1 backbone along with the NiV F and G encoding vectors, or with empty vector (pCAGGs). Culture supernatants were collected 36 hr post-transfection and filtered through a 0.45 μm filter and the pseudovirus preparations were normalized by p24 ELISA. The pseudovirus preparations were used to infect receptor positive and negative cells in triplicate wells and at 48 hr post infection, cells were lysed and assayed for luciferase reporter gene activity as described in the Methods. (B) Diagram of the panel of splice site mutants of the HeV G gene cloned into the pCAGGs vector. Putative splice donor sites are presented as black triangles and the splice acceptor site as grey squares. (C) Infection assay using pseudotyped virus particles prepared with HeV F along with (left to right) HeV G (wild-type) or each of the seven HeV G splice site mutants (SM1 - SM7); NiV G (wild-type); or empty vector (pCAGGs). Error bars indicate the standard error of the mean from triplicate wells.