FIG 8 .

A32 preferentially binds to cells that are CD4⁺ p24⁻ gag-pol mRNA⁻. (A and B) Primary CD4⁺ T cells infected with NL3.4 ADA GFP WT virus were stained with A32, followed by appropriate secondary Abs. Cells were then stained for phenotypic markers (see Materials and Methods) prior to detection of HIV-1 p24 and gag-pol mRNA by RNA-flow FISH. (A) Example of flow cytometry gating strategy based on A32 binding. (B) Quantification of the percentage of cells positive for HIV-1 p24 or gag-pol mRNA based on A32 binding. (C) Example of flow cytometry gating based on p24 and CD4 expression. (D) Quantification of the percentage of cells positive for A32 binding and gag-pol mRNA based on CD4 and p24 levels. (E) Quantification of the percentage of p24⁺ CD4⁺ cells among the cells positive for gag-pol mRNA. (F) Quantification of the percentage of cells that are CD4⁺ p24⁻, CD4⁺ p24⁺, or CD4⁻ p24⁺ among the cells recognized by A32. Error bars indicate means ± standard errors of the means of at least 4 independent experiments.