Mice given transplants of bm cotransduced with NUP98-HOXA10 and Flt3 succumb to AML. (A) Analysis of GFP (NUP98-HOXA10) and YFP (Flt3) expression in leukemic bm cells (2 representative profiles shown). (B) Southern blot analysis of genomic DNA harvested from the bm of leukemic mice. DNA digested with the LTR-specific NheI restriction endonuclease was probed with a GFP-specific probe confirming the integrity of the NUP98-HOXA10-GFP (4.5 kb) and Flt3-YFP (3.1 kb) proviruses. Lanes 1 and 2 show NUP98-HOXA10 + Flt3 transplant 4, bm, and spleen, respectively. Lanes 3 and 4 show NUP98-HOXA10 + Flt3 transplant 1, bm, and spleen, respectively. (C) Southern blot analysis of genomic DNA harvested from the spleen of leukemic mice. DNA was digested with a restriction endonuclease (HindIII) and probed with an Flt3-specific probe. Lane 1 shows NUP98-HOXA10 + Flt3 transplant 3; lane 2, NUP98-HOXA10 + Flt3 transplant 4; and lane 3, C57-negative control. (D) Immunophenotyping of bm cells. Top panel shows flow cytometric profiles on bm from a B6C3 control mouse; bottom panel shows bm from a representative NUP98-HOXA10 + Flt3 leukemic mouse. Expression of GFP/YFP is shown on the x-axis and the presence of the cell-surface antigens is shown on the y-axis. uns indicates unstained.