Fig. 3.
FISH analysis of vCWRHIVAPAP transduced and untransduced cells. CD34 were transduced with vCWRHIVAPAP at a functional MOI of 3 (particle MOI, 600) and placed in culture as described above. Suspension cells were harvested at designated time points for analysis. (A) A metaphase spread from an untransduced CD34 culture showing no hybridization of the vector-specific probe to cellular sequences. (B) Hybridization analysis of HIIC21, a G418-resistant clonal 293-based cell line derived after transduction with vCWRHIVASVN. This clone carries 1 copy of the vector genome per cell (28) and served as a positive control. (C, D, and E) Representative metaphases from vCWRHIVAPAP transduced CD34 cultures at 5 weeks posttransduction. Note that vector-specific signals are observed on both sister chromatids. Signals were not digitally enhanced in this analysis. (F) Representative interphase nuclei from a vCWRHIVAPAP transduced CD34 culture 3 weeks posttransduction. Three nuclei with three or more signals, two nuclei with two signals each, and three nuclei with no signals are seen.