Expression of neomycin phosphotransferase and β-gal in peripheral blood and bone marrow of in utero–transduced sheep at 40 months posttransduction
| Method of detection . | Sheep 690 . | Sheep 691 . | Sheep 692 . | Sheep 694 . | Sheep 697 . | Sheep 698 . | Sheep 703 . | Sheep 704 . | Sheep 705 . | |||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| PB . | BM . | PB . | BM . | PB . | BM . | PB . | BM . | PB . | BM . | PB . | BM . | PB . | BM . | PB . | BM . | PB . | BM . | |
| FDG FACS* | 5.1 | 3.2 | 2.3 | 2.6 | 0.4 | 1.7 | 0.4 | 3.9 | 0 | 2.5 | 2.5 | 2.7 | 2.5 | 1.9 | 1.3 | 1.2 | 1.2 | 5.3 |
| β-gal FACS† | 5.4 | 6.5 | 3.3 | 3.0 | 4.6 | 2.2 | 0.8 | 4.7 | 1.9 | 5.6 | 2.7 | 6.4 | 6.4 | 2.1 | 4.9 | 3.8 | 6.4 | 6.5 |
| NPT FACS† | 2.9 | 3.0 | 0 | 1.4 | 1.5 | 3.0 | 1.2 | 5.8 | 3.7 | 5.8 | 2.5 | 3.8 | 2.6 | 4.1 | 2.7 | 3.3 | 2.4 | 4.3 |
| Method of detection . | Sheep 690 . | Sheep 691 . | Sheep 692 . | Sheep 694 . | Sheep 697 . | Sheep 698 . | Sheep 703 . | Sheep 704 . | Sheep 705 . | |||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| PB . | BM . | PB . | BM . | PB . | BM . | PB . | BM . | PB . | BM . | PB . | BM . | PB . | BM . | PB . | BM . | PB . | BM . | |
| FDG FACS* | 5.1 | 3.2 | 2.3 | 2.6 | 0.4 | 1.7 | 0.4 | 3.9 | 0 | 2.5 | 2.5 | 2.7 | 2.5 | 1.9 | 1.3 | 1.2 | 1.2 | 5.3 |
| β-gal FACS† | 5.4 | 6.5 | 3.3 | 3.0 | 4.6 | 2.2 | 0.8 | 4.7 | 1.9 | 5.6 | 2.7 | 6.4 | 6.4 | 2.1 | 4.9 | 3.8 | 6.4 | 6.5 |
| NPT FACS† | 2.9 | 3.0 | 0 | 1.4 | 1.5 | 3.0 | 1.2 | 5.8 | 3.7 | 5.8 | 2.5 | 3.8 | 2.6 | 4.1 | 2.7 | 3.3 | 2.4 | 4.3 |
Detection of β-gal activity by fluorescence-activated cell sorter (FACS) analysis using fluorescein di-β-D-galactopyranoside (FDG) as substrate from in utero–transduced sheep. Values represent the percentage of positive cells per 50 000 viable white cells after subtracting the values obtained with a control sheep.
Data obtained from FACS analysis of peripheral blood (PB) and bone marrow (BM) white cells from in utero–transduced sheep using antibodies to β-gal and neomycin phosphotransferase (NPT). Values represent the percentage of positive cells per 20 000 white cells after subtracting the minimal (< 0.2%) values obtained with a control sheep using the same gates. Expression of β-gal and NPT was also confirmed by performing immunofluorescence microscopy, and similar values were obtained, with no significant staining seen in any of the control sheep tested (data not shown). For FACS histograms representative of the staining obtained with these antibodies, see references 19, 20, and 54.