rVSV-EnvG<sub>4</sub>-G<sub>6</sub> characterization.

<p>(a) After a 24 hr infection, total infected Vero cell lysates were collected and proteins were separated by SDS polyacrylamide gel electrophoresis (SDS-PAGE). Blot was probed with anti-VSV-N and anti-VSV-G<sub>IN</sub> CT, which does not recognize G<sub>NJ</sub>. IN: rVSV-EnvG<sub>4</sub>-G<sub>6</sub><sup>IN</sup>; NJ: rVSV-EnvG<sub>4</sub>-G<sub>6</sub><sup>NJ</sup>. (b-d) Sucrose-gradient purified rVSV-EnvG<sub>4</sub>-G<sub>6</sub> particles were separated by SDS-PAGE. (b) Blots of 10<sup>6</sup> pfu of rVSV-EnvG<sub>4</sub>-G<sub>6</sub><sup>IN</sup> and 2.5×10<sup>6</sup> pfu rVSV-EnvG<sub>4</sub>-G<sub>6</sub><sup>NJ</sup> vectors were probed separately for EnvG using anti-gp120. (c) Blot of 10<sup>6</sup> pfu rVSV-EnvG<sub>4</sub>-G<sub>6</sub><sup>IN</sup> was probed with anti-VSV-G<sup>IN</sup> CT. (d) Blot of 10<sup>6</sup> pfu rVSV-EnvG<sub>4</sub>-G<sub>6</sub><sup>IN</sup> and rVSV-EnvG<sub>4</sub>-G<sub>6</sub><sup>NJ</sup> was probed with anti-VSV-G. (e) Replication kinetics of rVSV-EnvG<sub>4</sub>-G<sub>6</sub> and rVSV-G<sub>4</sub> viruses in Vero cells. Vero cells were infected in 6-well plates at an MOI of 0.1. At various intervals post infection, supernatant was collected from duplicate wells and virus was titrated. Plaques were counted manually after cell fixation.</p>