Stably expressed APOBEC3H forms a barrier for cross-species transmission of simian immunodeficiency virus of chimpanzee to humans
Fig 5
Inhibition of SIVcpz by hA3H haplotypes with stable protein expression.
(a) SIVcpzPttMB897WT or SIVcpzPttMB897Δvif reporter constructs were co-transfected with increasing amounts of hA3H hapI or hapII expression plasmids (5, 10, 30, 100 or 200 ng), PTR600 empty vector was used to bring the total transfected plasmid DNA to 600 ng and also used as control (vector). Two days post-transfection, normalized amounts of viruses were used to infect 293T cells. The nanoluciferase (relative light units-RLU) was measured two days post-infection. (b) Cell lysates from (a) were used to detect the expression of A3s, SIVcpz capsid (p24), or SIVcpz Vif by anti-HA, anti-p24, or anti-Vif antibody, respectively. Tubulin served as a loading control. (c) SIVcpzPttMB897WT or SIVcpzPttMB897Δvif reporter constructs were co-transfected but with expression plasmids for different A3H haplotypes and splice variants. (d) Cell lysates from (c) were used to detect the expression of A3s by anti-HA antibody. Tubulin served as a loading control. (e) SupT11-vetor-hCCR5 or SupT11-hA3H hapII-hCCR5 cells were infected with 1 ng RT activity of SIVcpzPtsTAN1, SIVcpzPttMB897 or SIVcpzPttGab1, respectively, and culture supernatants were collected each second day and quantified by the RT assay.