In all infectivity experiments cells were infected with HIV-1 at a multiplicity of infection (MOI) of 0.1.
For infectivity experiments, mice were orally inoculated with saline or 109S.
M.D.S. worked together with R.S.H. on all aspects of these studies and did most of the experiments; M.B.B. and M.L. established the TK mutation assay and the in vitro DNA deamination assays, respectively; J.L. performed the virus infectivity experiments.
All viruses used in infectivity experiments were plaque-assayed in parallel.
The 1∶1 mixture was used to detect possible infectious bottlenecks [11] in the infectivity experiments.
This conclusion was further supported by the results obtained from Ae. aegypti infectivity experiments.
Infectivity experiments were performed essentially as for the LD50 experiments except a single dose (averaging 4×108 cfu) was used in all cases.
Single cycle infectivity experiments confirmed that inhibition of Vif-deficient HIV-1 by A3G was maintained in the presence of these mutations and that all proteins were expressed in similar amounts as determined by immunoblotting (Figure 2A).
In spite of comparable results obtained with non-fractionated samples, in some infectivity experiments ASL was size-fractionated by gravity sedimentation (after suspension in water), so as to be better tolerated by cells.
Tenfold serial dilutions were prepared from supernatants harvested from individual wells of the infectivity experiments.
