Engraftment of donor-derived cells in bone marrow and spleen of primary BMT recipients preconditioned with 1 Gy TBI.

Surviving animals (depicted in Figure 3) were killed 120 days after transplantation. Genomic DNA samples isolated from bone marrow and spleen were subjected to quantitative Southern analysis as described in “Materials and methods.” (A) Representative Southern image of selected spleen samples. A DHFR probe was used to quantitate total DNA (En DHFR) and donor-derived DHFR transgenic DNA (Tg04 DHFR). An APP probe was used to quantitate donor-derived normal cells (APP), and a Y probe was used to quantitate the percentage of host cells (Y). (B) Levels of engrafted donor-derived cells. Signals from the Southern image were quantified using a PhosphorImager. Ratios of APP/En DHFR and Tg DHFR/En DHFR were used to calculate the percentage of donor-derived normal and Tg04 cells, respectively, from a standard curve. Values are presented as mean ± SD of 5 to 9 samples.