Patient-derived Vifs were cloned into an expression vector and co-transfected with NL4-3ΔVif and WT A3G (A) or A3G-H186R (B) expression plasmids and infectivity of the produced viruses was tested on TZM-bl reporter cells. NS; not significant, unpaired t test. 293T cell lysates were analysed by western blot and probed for HA, Vif and GAPDH serves as a loading control. Vif variants derived from homozygotes WT A3G are represented by black bars while Vifs derived from A3G-H186R homozygous carriers are represented by red bars. Error bars represent standard deviations from triplicate transfections. (C) Dot plot comparing the infectivity levels in the presence of WT A3G or A3G-H186R, P < 0001, paired t test. (D) Comparison of infectivities in the presence of WT A3G and A3G-H186R in the absence of Vif (also shown in 2A). P = 0.0007, unpaired t test. Error bars represent standard deviations from triplicate transfections.