Fig. 3

Effect of viral RNA length on reverse transcription and uncoating. a Schematic representation of pCSGW and pHIVLacZ plasmids highlighting key positions in bp. b, c 293T cells were infected with either WT GFP-HIV (green) or LacZ-HIV (red) VLP. At the indicated times post-infection, the level of cDNA corresponding to b early or c late reverse transcription products were analysed by qPCR. d CsA washout assay. OMK cells were infected with GFP-HIV or LacZ-HIV VLP in the presence of CsA. CsA was removed from each sample at the indicated times. Infection was measured after 72 h and plotted relative to that observed when CsA was removed 4 h.p.i. e Modified fate-of-CA assay. 293T cells were infected with GFP-HIV or LacZ-HIV VLP. Cells were lysed 4 h.p.i. and lysate separated in a 10–50 % (w/v) sucrose gradient. The gel shows a representative image from 3 biological repeats. The CA content in each fractions was detected by immunoblotting, quantified and the percentage of CA relative to the total cell lysate (input) was calculated and plotted. Each graphical panel shows the mean and SEM of at least 3 independent experiments