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. 2002 Jun 11;99(12):8295–8300. doi: 10.1073/pnas.122131099

Figure 4.

Figure 4

DNA analysis of vector integrations and provirus copy number. (A) Southern blot of genomic DNA from expanded human myeloid cells (M) and three B cell lines derived from mouse 3D were digested with BclI or NsiI and hybridized to probe P1 (Fig. 1). B cell lines 1 and 3 have identical patterns, indicating that they represent the same clonal population. (B) Southern blots of genomic DNA from expanded human myeloid cells (M) and 12 B cell lines isolated from mouse 4B. Junction fragments (Fig. 5) found to be present in both lymphoid and myeloid populations are indicated by asterisks. (C) Determination of vector copy number in expanded human GFP+ and GFP− myeloid cells from mice 4B, 4C, and 4D. BclI-digested DNA was hybridized to probe P2 (Fig. 1), identifying a 3.5-kb internal fragment in all integrated proviruses. Dilution standards are mixtures of DNA from diploid human fibroblasts containing a single FV vector provirus with DNA from normal human lymphocytes. Copy numbers (proviruses/diploid genome) determined by PhosphoImager quantitation are indicated below each sample. Correction for DNA loading was based on reprobing the same blot with a fragment of the human COL1A1 gene.