Fig. 4

HIV-1 Gag-iNanoLuc vector recapitulates inhibition of HIV-1 particle release. a NanoLuc activity in lysates and supernatants from HEK293T cells transfected in 12-well plates with 1.0μg of either pNL4-3 Gag-iNanoLuc WT, pNL4-3 Gag-iNanoLuc PTAP- or pNL4-3 Gag-iNanoLuc delVpu and either 0, 0.1μg or 0.2μg HA-tetherin (BST2). NanoLuc activity in lysates and supernatants was measured at 48 h post-transfection and normalized to that of pNL4-3 Gag-iNanoLuc WT. b Western blot analysis of HIV-1 Gag (upper) in lysates and supernatant from a and HA-tetherin (lower) in cell lysates. c NanoLuc activity in lysates and supernatants from HEK293T cells transfected with 1.0μg of pNL4-3 Gag-iNanoLuc and treated with either 5μM or 10μM AME or vehicle at 48 h post-transfection. AME treatment was done at 24 h post-transfection following replacement of the growth media. d Western blot analysis of HIV-1 Gag in lysates and supernatant from c. Error bars ± SD, n = 3 independent experiments for a and c. *, P < 0.05 by one-way ANOVA in panels A and C compared to WT pNL4-3 NanoLuc activity and the 0 AME control NanoLuc activity, respectively