Fig. 2.
Fig. 2. Cbfβ is expressed at uniform levels in most hematopoietic tissues ofCbfb+/GFP mice, but shows differential expression in hematopoietic lineages isolated from bone marrow. / Cells were isolated from adult (6-month-old)Cbfb+/GFP and Cbfb+/+mice and analyzed by FACS. Representative histograms show the distribution of cells with respect to GFP fluorescence. Dashed line (– –) represents Cbfb+/+ autofluorescence; solid line (___) represents fluorescence fromCbfb+/GFP animals. (A) Expression of Cbfβ-GFP in the indicated tissues. (B) Cells were isolated from bone marrow (BM) of adult Cbfb+/+andCbfb+/GFP mice, enucleated cells were lysed, and the remaining cells were analyzed for GFP expression (left panel). Bone marrow cells were also stained with PE-conjugated antibodies against Mac1, GR1, or GPIIb/IIIa. The positively stained cells were gated and analyzed for GFP expression. (C) Cells were isolated from bone marrow of adult Cbfb+/+andCbfb+/GFP mice, lysed in ACK lysing buffer, and stained with APC-conjugated anti–c-kit and PE-conjugated anti-TER119. A representative contour plot (c-kit–APC versus Ter119-PE) is shown. Cells from both wild-type and heterozygous mice were gated into the following populations and analyzed for GFP expression: c-kit+/Ter119− (R2 = 2.3%), c-kit+/Ter119+ (R3 = 0.2%), c-kit−/Ter119lo (R4 = 0.6%), c-kit−/Ter119hi (R5 = 2.4%). (D) Nucleated cells from Cbfb+/GFP andCbfb+/+ bone marrow were separated into Ter119-enriched and Ter119-depleted populations by magnetic sorting using Ter119 microbeads. Cells from each population were analyzed by Western blot: Ter119-depleted cells fromCbfb+/GFP (lane 1) and Cbfb+/+(lane 2) bone marrow, and the Ter119-enriched population fromCbfb+/+ bone marrow (lane 3). MEN1 indicates multiple endocrine neoplasia 1; *, nonspecific bands. (E) Cells were isolated from bone marrow of adultCbfb+/+ and Cbfb+/GFPmice and stained with APC-conjugated anti-B220 and the markers indicated above each histogram. The particular B-cell population being examined is also indicated in the upper right hand corner of the graphs. Cells from wild-type and heterozygotes were gated appropriately and analyzed for GFP expression.

Cbfβ is expressed at uniform levels in most hematopoietic tissues ofCbfb+/GFP mice, but shows differential expression in hematopoietic lineages isolated from bone marrow.

Cells were isolated from adult (6-month-old)Cbfb+/GFP and Cbfb+/+mice and analyzed by FACS. Representative histograms show the distribution of cells with respect to GFP fluorescence. Dashed line (– –) represents Cbfb+/+ autofluorescence; solid line (___) represents fluorescence fromCbfb+/GFP animals. (A) Expression of Cbfβ-GFP in the indicated tissues. (B) Cells were isolated from bone marrow (BM) of adult Cbfb+/+andCbfb+/GFP mice, enucleated cells were lysed, and the remaining cells were analyzed for GFP expression (left panel). Bone marrow cells were also stained with PE-conjugated antibodies against Mac1, GR1, or GPIIb/IIIa. The positively stained cells were gated and analyzed for GFP expression. (C) Cells were isolated from bone marrow of adult Cbfb+/+andCbfb+/GFP mice, lysed in ACK lysing buffer, and stained with APC-conjugated anti–c-kit and PE-conjugated anti-TER119. A representative contour plot (c-kit–APC versus Ter119-PE) is shown. Cells from both wild-type and heterozygous mice were gated into the following populations and analyzed for GFP expression: c-kit+/Ter119− (R2 = 2.3%), c-kit+/Ter119+ (R3 = 0.2%), c-kit−/Ter119lo (R4 = 0.6%), c-kit−/Ter119hi (R5 = 2.4%). (D) Nucleated cells from Cbfb+/GFP andCbfb+/+ bone marrow were separated into Ter119-enriched and Ter119-depleted populations by magnetic sorting using Ter119 microbeads. Cells from each population were analyzed by Western blot: Ter119-depleted cells fromCbfb+/GFP (lane 1) and Cbfb+/+(lane 2) bone marrow, and the Ter119-enriched population fromCbfb+/+ bone marrow (lane 3). MEN1 indicates multiple endocrine neoplasia 1; *, nonspecific bands. (E) Cells were isolated from bone marrow of adultCbfb+/+ and Cbfb+/GFPmice and stained with APC-conjugated anti-B220 and the markers indicated above each histogram. The particular B-cell population being examined is also indicated in the upper right hand corner of the graphs. Cells from wild-type and heterozygotes were gated appropriately and analyzed for GFP expression.

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