Figure 5.

Stabilized and packaged A3H haplotype III is unable to restrict HIV-1. (A) Single-cycle infectivity assays performed with vif-deficient HIV-1 virus produced in the presence of the indicated wild-type A3H haplotypes or chimeras. Shown is the average of three biological replicates with error bars indicating the standard deviation of the mean. * p < 0.05, ** p < 0.01, *** p < 0.001, ns = not significant (p > 0.05), unpaired t test. (B) Immunoblot of viral supernatant (Virus, top) and whole cell lysate (Cells, bottom) of virus producing cells as in (A) using anti-A3H to assess levels of A3H packaged in virus or expressed in cells. Anti-p24 and anti-Lamin B1 were used as loading controls for virus and cell lysate, respectively. (C) Single-cycle infectivity assays performed with vif-deficient HIV-1 virus produced in the presence of the indicated A3H chimeras. Shown is the average of three biological replicates with error bars indicating the standard deviation of the mean. ** p < 0.01, unpaired t test. E56A is a mutation at the catalytic active site of hap II (D) Immunoblot of indicated A3H chimeras expressed in 293T cells. Western blotting of whole cell lysates was performed using anti-A3H and anti-Lamin B1 as a loading control.