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. 2023 Aug 9;14(4):e00782-23. doi: 10.1128/mbio.00782-23

Fig 1.

Fig 1

Endogenous A3H protein does not inhibit HIV-1 in THP-1 cells. (A) A3H haplotypes in THP-1 cells. The indicated positions are key amino acid residues that determine the expression of unstable (hapI) or stable (hapII) A3H protein. (B) Schematic of the susceptibility of HIV-1 Vif mutants to antiviral activity by stable A3H haplotypes. Key amino acid residues of Vif that determine the susceptibility of HIV-1 IIIB to restriction by stable A3H haplotypes. −, full resistance; +, partial resistance; +++, sensitivity. (C) Schematic depiction of the pseudo-single cycle infectivity assay. For details, see the main text and the “Pseudo-single cycle infectivity assays” section in Materials and Methods. (D) Representative infectivity of HIV-1 mutants with hyper- and hypo-functional Vif produced from THP-1 cells. Top panels show the infectivity of hyper-Vif, hypo-Vif, IIIB Vif, and Vif-deficient HIV-1 mutants produced in THP-1 cells. The amounts of produced viruses used to infect TZM-bl cells were normalized to p24 levels. Each bar shows the average of four independent experiments with standard deviation (SD). Data are represented as relative infectivity compared to hyper-Vif HIV-1. Statistical significance was determined using the two-sided paired t test. *P < 0.05 compared with the infectivity of hyper-Vif HIV-1. The bottom panels are representative western blots of three independent experiments. The levels of viral and cellular proteins in viral particles and whole-cell lysates are shown. p24 and HSP90 were used as loading controls. (E) G-to-A mutations. Average number of G-to-A mutations in the 564 bp pol gene after infection with hyper-Vif, hypo-Vif, IIIB Vif, or Vif-deficient HIV-1 produced from THP-1. Each bar depicts the average of three independent experiments with SD. (F) G-to-A mutation profile. Dinucleotide sequence contexts of G-to-A mutations in the 564 bp pol gene after infection with the indicated viruses produced from indicated cell lines. Each vertical line indicates the location of the dinucleotide sequence contexts described in the legend within the 564 bp amplicon (horizontal line).