NSG-engrafting AML cells reconstitute secondary recipient. (A) NSG-engrafting AML MNCs were injected in NOD/SCID β2^−/− and NOD/SCID IL2Rγc^null mice. Engrafted BM cells from NOD/SCID IL2Rγc^null-transplanted mice were injected in secondary irradiated NOD/SCID β2^−/− and NOD/SCID IL2Rγc^null mice. Mice were analyzed 8-weeks after transplantation by flow cytometry. (B) Representative flow cytometry plots of human engraftment in primary and secondary recipients after NSG-engrafting AML injection (AML#23). (C) Phenotype analysis of primary (left panel) and secondary (central panel) BM grafts of NSG-engrafting AML (AML#23)-transplanted mice. Left panel corresponds to primary spleen engrafted cells. Myeloid (CD33 and CD13 inside CD45-positive cells), B (CD19 inside CD45-positive cells), NK (CD2 and CD56 inside CD45-positive cells), T cells (CD3, and CD4/CD8 inside CD3-positive cells) are represented. (D) Analysis of primary (left panel) and secondary (right panel) graft of AML (AML#21)- and T-ALL–transplanted mice based on the myeloid (CD33 and CD13 inside CD45-positive cells), B (CD19 inside CD45-positive cells) and T (CD3, and CD2/CD7 inside CD45-positive cells) surface markers.