Fig. 1
Generation of HIV-1 Gag-NanoLuc vectors. a A schematic representation of the HIV-1 Gag polyprotein precursor indicating different domains (black rectangles) and PR cleavage sites (white lines). The PR cleavage site between the MA and CA domains of Gag (SQNYPIVQ) is shown with the dashed line. The NanoLuc gene is represented by the red rectangle. In the HIV-1 Gag-iNanoLuc vector, the cleavage site motif is duplicated to flank the NanoLuc gene on both sides. Synonymous changes made in the duplicated coding sequence are represented in bold and italics. Maturation products of the different vectors are indicated on the right, with approximate molecular masses indicated. b Western blot analysis of HIV-1 Gag in lysates and supernatants from HEK293T cells transfected with the indicated HIV-1 vectors at 48 h post-transfection. Expected molecular masses of the Gag products are shown on the left; molecular mass markers are shown on the right. c NanoLuc activity of lysates and supernatants from HEK293T cells from (b) represented as Log10[RLUs]. d Relative virus release efficiency in HEK293T cells from (b). Error bars ± SD, n = 3 independent experiments for both c and d. *, P < 0.05 by one-way ANOVA compared to WT virus release efficiency