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Table I.

B cell–specific Transgenic Expression of Bcl2 Rescues B Cell Numbers and Maturation Phenotypes in Bone Marrow and Spleen of BOB.1/OBF.1-deficient Mice




Wild type

Bcl2 tg

BOB.1/OBF.1 ko

Bcl2 tg × BOB.1/OBF.1 ko
 Absolute cell number × 107    
Spleen     
Total cellularity 9.0 ± 0.7 22.3 ± 2.2 6.2 ± 1.2 21.7 ± 1.6 
B220+ 5.5 ± 0.9 16.8 ± 1.3 2.3 ± 0.3 14.5 ± 2.5 
T1 1.8 ± 0.3 4.6 ± 0.5 1.1 ± 0.1 6.1 ± 0.6 
T2 1.9 ± 0.3 5.6 ± 0.9 0.6 ± 0.1 3.7 ± 1.3 
1.8 ± 0.3 5.7 ± 0.7 0.6 ± 0.1 3.8 ± 1.1 
Annexin V+/B220 0.46 ± 0.24 0.89 ± 0.29 0.25 ± 0.08 1.22 ± 0.59 
Bone marrow     
Total cellularity 2.3 ± 0.4 3.3 ± 1.1 2.0 ± 0.7 2.7 ± 0.9 
Pro/Pre B 0.54 ± 0.06 0.61 ± 0.08 0.64 ± 0.15 0.64 ± 0.15 
Immature 0.32 ± 0.08 0.89 ± 0.14 0.33 ± 0.14 0.92 ± 0.09 
Transitional 0.12 ± 0.02 0.31 ± 0.11 0.04 ± 0.01 0.16 ± 0.05 
Recirculating 0.23 ± 0.06 0.57 ± 0.16 0.06 ± 0.04 0.26 ± 0.06 
 % % % % 
Spleen     
B220+ 61.9 ± 9.7 75.3 ± 5.7 36.7 ± 5.2 66.6 ± 11.4 
T1 20.4 ± 2.9 20.8 ± 2.2 16.9 ± 1.6 27.9 ± 2.7 
T2 21.0 ± 5.8 24.9 ± 3.9 10.0 ± 1.1 17.1 ± 6.1 
19.4 ± 3.6 25.4 ± 3.3 9.1 ± 2.0 17.6 ± 4.9 
Annexin V+/B220 8.3 ± 4.3 5.3 ± 1.7 10.7 ± 3.4 8.4 ± 4.1 
Bone marrow     
Pro/Pre B 23.3 ± 2.5 18.6 ± 2.5 32.2 ± 7.3 23.7 ± 5.6 
Immature 14.0 ± 3.4 26.9 ± 4.3 16.4 ± 7.2 34.1 ± 3.2 
Transitional 5.1 ± 0.6 9.5 ± 3.4 1.8 ± 0.5 5.8 ± 1.8 
Recirculating
 
10.0 ± 2.7
 
17.2 ± 5.0
 
3.2 ± 2.1
 
9.5 ± 2.2
 



Wild type

Bcl2 tg

BOB.1/OBF.1 ko

Bcl2 tg × BOB.1/OBF.1 ko
 Absolute cell number × 107    
Spleen     
Total cellularity 9.0 ± 0.7 22.3 ± 2.2 6.2 ± 1.2 21.7 ± 1.6 
B220+ 5.5 ± 0.9 16.8 ± 1.3 2.3 ± 0.3 14.5 ± 2.5 
T1 1.8 ± 0.3 4.6 ± 0.5 1.1 ± 0.1 6.1 ± 0.6 
T2 1.9 ± 0.3 5.6 ± 0.9 0.6 ± 0.1 3.7 ± 1.3 
1.8 ± 0.3 5.7 ± 0.7 0.6 ± 0.1 3.8 ± 1.1 
Annexin V+/B220 0.46 ± 0.24 0.89 ± 0.29 0.25 ± 0.08 1.22 ± 0.59 
Bone marrow     
Total cellularity 2.3 ± 0.4 3.3 ± 1.1 2.0 ± 0.7 2.7 ± 0.9 
Pro/Pre B 0.54 ± 0.06 0.61 ± 0.08 0.64 ± 0.15 0.64 ± 0.15 
Immature 0.32 ± 0.08 0.89 ± 0.14 0.33 ± 0.14 0.92 ± 0.09 
Transitional 0.12 ± 0.02 0.31 ± 0.11 0.04 ± 0.01 0.16 ± 0.05 
Recirculating 0.23 ± 0.06 0.57 ± 0.16 0.06 ± 0.04 0.26 ± 0.06 
 % % % % 
Spleen     
B220+ 61.9 ± 9.7 75.3 ± 5.7 36.7 ± 5.2 66.6 ± 11.4 
T1 20.4 ± 2.9 20.8 ± 2.2 16.9 ± 1.6 27.9 ± 2.7 
T2 21.0 ± 5.8 24.9 ± 3.9 10.0 ± 1.1 17.1 ± 6.1 
19.4 ± 3.6 25.4 ± 3.3 9.1 ± 2.0 17.6 ± 4.9 
Annexin V+/B220 8.3 ± 4.3 5.3 ± 1.7 10.7 ± 3.4 8.4 ± 4.1 
Bone marrow     
Pro/Pre B 23.3 ± 2.5 18.6 ± 2.5 32.2 ± 7.3 23.7 ± 5.6 
Immature 14.0 ± 3.4 26.9 ± 4.3 16.4 ± 7.2 34.1 ± 3.2 
Transitional 5.1 ± 0.6 9.5 ± 3.4 1.8 ± 0.5 5.8 ± 1.8 
Recirculating
 
10.0 ± 2.7
 
17.2 ± 5.0
 
3.2 ± 2.1
 
9.5 ± 2.2
 

The total cell numbers per spleen or femur from animals at 8–9 wk of age of each genotype were determined. Before analyses splenic cells were purified by Ficoll gradient. Bone marrow cells were stained with a combination of anti-B220 and anti-IgM antibodies to discriminate between pro/pre–B cells, immature, transitional and recirculating B cells. Splenocytes were stained with a mixture of anti-B220, anti-IgM, and anti-IgD antibodies to distinguish between the IgMhighIgDlow (T1), IgMhighIgDhigh (T2), and IgMlowIgDhigh (M) populations and to determine the overall B cell numbers. Additionally, splenocytes were stained with a mixture of anti-B220 and anti–Annexin V antibodies. Mean percentages as well as absolute numbers for B cells of each compartment with standard deviation out of six to eight animals of each genotype are given.

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