A RT I C L E Figure 1. Phenotypic analysis of GFP+ cells in various tissues after transplantation of ICN1-overexpressing BM cells. (a) Single-cell sus- pensions were prepared from blood and BM at 2 wk and from blood, lung, and liver at 8 wk after reconstitution with retrovirally transduced BM. GFP+ and GFP cells were stained with CD4 and CD8 antibodies. Infiltration by GFP+CD4+8+ and GFP+CD4 8+ malignant cells was analyzed in nonlymphoid organs such as lung and liver. Numbers indicate the percentage of cells within each quadrant. Data are representative of three independent experi- ments. (b) BrdU–labeling kinetics: continuous (top) and pulse (bottom) BrdU labeling of lymphocytes subsets. BrdU was administered to control mice and mice at 2 or 8 wk after BM transplantation with ICN1-overexpressing cells. The percentage of BrdU-labeled cells was analyzed by flow cytometry as previously described (13). (c) CFSE labeling: normal CD4+8+ thymocytes and ICN1-overexpressing CD4+8+ splenocytes were labeled with CFSE and injected into nu/nu mice. Dilution of CFSE label was analyzed by FACS analysis. (d) BrdU–labeling kinetics: continuous (top) and pulse (bottom) BrdU labeling of ICN1-overexpressing TCR- CD4 8+ splenocytes versus control normal CD4 8+ thymocytes. (e) TCR- expression by various lympho- cyte subsets in ICN1-transplanted mice at 2 wk after BMT, as well as control mice. TCR- staining was performed with the pan–TCR- antibody H57. JEM VOL. 205, November 24, 2008 2853