FIG. 6.

FACS analysis of lacZ gene expression in erythroid- and myeloid-differentiated CD34⁺ primary human bone marrow cells transduced with recombinant AAV- and B19-lacZ vectors. Primary human bone marrow-derived CD34⁺ cells were allowed to undergo differentiation into erythroid or myeloid lineages with the use of respective cytokine combinations for 10 days in vitro, as described in Materials and Methods. Following differentiation, cells were either mock infected or infected with either 1 × 10⁵ particles of AAV-lacZ vector per cell or 2 × 10² particles of B19-lacZ vector per cell under identical conditions. Forty-eight hours postinfection, cells were harvested and stained with either PE-conjugated glycophorin A antibody (stippled bars) or PE-conjugated CD33 antibody (solid bars) and analyzed for lacZ gene expression in erythroid and myeloid populations by using fluorescent β-Gal product, as described in the legend to Fig. 4. Upper-row panels at the top are FACS dot plots from a representative experiment indicating cells positive for lacZ (FL1-H) and glycophorin A (FL2-H). Lower-row panels at the top represent cells expressing lacZ (FL1-H) and CD33 (FL2-H). The results of dual positive cells from three separate experiments are plotted in the bar graph.