Characterization of the rAAV-LacZ DNA in FACS-purified mature CD3⁺ T cells derived from transduced CD34⁺CD2⁻ cells. (A) Cells cultured for 28 days expressed surface CD3, CD4, and CD8 and were stimulated with IL-2 and PHA as described in the Materials and Methods. CD3⁺CD4⁺ and CD3⁺CD8⁺ cells were sorted by flow cytometry using the depicted gates after quadrants were established with matched isotype control antibodies. (B) PCR analysis of high molecular weight DNA extracted from purified mature single-positive T cells. PCR and Southern blot analysis for LacZ transgene was performed as described in Fig 5. Primers for β-actin were used as a positive control. Intervening wells between the last two lanes have been spliced out for clarity.