Intermediate HPCs derived from the bone marrow of HIV-infected NSG-BLT mice are productively infected as shown by HIV-GAG expression. (A) Schematic of experimental design for infecting humanized NSG-BLT mice and isolating CD34+ HPCs from their bone marrow. (B) Peripheral blood mononuclear cells from infected mice were analyzed by PCR for the presence of full-length viral DNA relative to the number of copies of human β-globin to determine the percent of infected cells. Each mark represents the average of 3 reactions for each mouse; the total data represent the pooled mean ± SEM of all reactions from all mice. Cells isolated from the murine bone marrow were analyzed for the presence of human intermediate HPC markers in conjunction with HIV-GAG expression. (C) Cells derived from uninfected mice. (D) Cells derived from mice exposed to HIV_NL4-3. (E) Representative flow cytometric analysis of CD34+ purity after sorting from BLT bone marrow. (Left) Isotype control; (right) CD34 staining at 98.5% purity. (F) Human CD34+ HSC derived from BLT bone marrow harbor full-length HIV DNA when analyzed by quantitative PCR. Each mark indicates the average of triplicate assays in a single mouse; the total data represent the pooled mean ± SEM of all reactions from all mice. The dashed line indicates the limit of detection of the assay. JRCSF, the patient code used to define the particular strain of HIV.