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Fig. 1. The Scl 3' enhancer construct targets lacZ expression in transgenic mice to long-term reconstituting haematopoietic stem cells from adult bone marrow and foetal liver. (A) FACS analysis of cell suspensions from non transgenic and transgenic adult bone marrow for the expression of lacZ by FDG staining on gated live cells (PI-negative cells). Histograms for FDG expression in one representative experiment are shown. Cells included in the sorted windows were designated as FDG– and FDG+. (B) Ethidium bromide-stained gels of peripheral blood PCR analysis for the presence of the transgene (lacZ) together with an internal control (myogenin). PCR products are shown from a representative experiment performed 5 months post-transplantation with bone marrow cells. Lanes correspond to individual animals transplanted with the indicated number of FDG+ or FDG– cells. Peripheral blood from transgenic and non-transgenic mice was mixed to provide 100%, 10%, 1% and 0% lacZ controls, allowing semi-quantitative evaluation of reconstitution levels. (C) Multilineage analysis in one recipient mouse 12 months post-transplant with FDG+ bone marrow cells. (D) FACS analysis of cell suspensions from non transgenic and transgenic foetal liver for the expression of lacZ by FDG staining on gated live cells (PI-negative cells). Histograms for FDG expression in one representative experiment from E12 liver cells are shown. (E) lacZ-PCR analysis of peripheral blood 4 months post-transplantation with foetal liver cells. (F) Multilineage analysis of a recipient mouse 6 months post transplant with foetal liver FDG+ cells. Bb, B lymphocytes B220+ from bone marrow; Bs, B lymphocytes B220+ from spleen; Bl, blood; BM, bone marrow; Erb, erythocytes Ter119+ from bone marrow; Grb, granulocytes Gr-1+ from bone marrow; LN, lymph nodes; Mø*, peritoneal cells; Møb, macrophages CD11b+ from bone marrow; Møs, macrophages CD11b+ from spleen; Sp, spleen; Th, thymus; Ts, T lymphocytes CD4+CD8+ from spleen; TFL, total foetal liver. The 2262 transgenic line was used in the experiments shown.





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