Fig. 6.

Effects of mutations affecting the IN/H4 interaction during early steps of viral replication. HEK-293T cells were transduced with VSV-G pseudotyped lentiviruses encoding either WT IN or the R231A/H/G or D253H IN mutants or the catalytically inactive class I D116A mutant with or without AZT 1 µM. Viral replication was quantified based on eGFP fluorescence measured by FACS 48 h post-transduction. The data shown in (a) are expressed as the percentage of eGFP-positive cells at a MOI of 1. The replication steps affected by the mutations were determined by measuring the amounts of the different viral DNA species produced using qPCR. Levels of total viral DNA, integrated DNA and 2-LTR circles shown respectively in (b) were monitored between 0 and 72 h post-transduction to check for potential defects at the steps of reverse transcription, integration and nuclear import of the preintegration complex, respectively. The data are represented as the mean of at least three independent experiments ± standard deviation. The p-values were calculated by Student’s t-test and are shown as *p < 0.05 and **p < 0.005 to represent the probability of obtaining significant differences compared with the WT data