Basic Information
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Targeting strategy
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Gene targeting strategy for B-hLTA/hTNFA/hLTB mice.
The targeted mouse Lta, Tnfα and Ltb whole genomic sequence was replaced by human LTA, TNFα and LTB whole genomic sequence in B-hLTA/hTNFA/hLTB mice.
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mRNA expression analysis
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Strain specific analysis of LTA, TNFA, LTB gene expression in homozygous B-hLTA/hTNFA/hLTB mice by RT-PCR. Mouse Lta, Tnfa and Ltb mRNA were detectable in splenocytes of wild-type (+/+) mice. Human LTA, TNFα and LTB mRNA were detectable only in homozygous B-hLTA/hTNFA/hLTB mice (H/H), but not in wild-type mice.
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Protein expression analysis
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Strain specific LTA expression analysis in homozygous B-hLTA/hTNFA/hLTB mice by ELISA. Serum was collected from wild-type mice (+/+) and homozygous B-hLTA/hTNFA/hLTB mice (H/H) stimulated with LPS in vivo, and analyzed by ELISA with species-specific LTA ELISA kit. Mouse LTA was both detectable in wild-type mice and B-hLTA/hTNFA/hLTB mice, as the antibody is crossly reactive with human LTA.
Strain specific TNFA expression analysis in homozygous B-hLTA/hTNFA/hLTB mice by ELISA. Serum was collected from wild-type mice (+/+) and homozygous B-hLTA/hTNFA/hLTB mice (H/H) stimulated with LPS in vivo, and analyzed by ELISA with species-specific TNFα ELISA kit. Mouse TNFα was detectable in wild-type mice. Human TNFα was detectable in homozygous B-hLTA/hTNFA/hLTB mice.
Strain specific LTB expression analysis in homozygous B-hLTA/hTNFA/hLTB mice by western blot. Spleen and thymus were collected from wild-type C57BL/6 mice (+/+) and homozygous B-hLTA/hTNFA/hLTB mice (H/H), and analyzed by western blot with anti-LTB antibody. Mouse LTB was detectable both in wild-type mice and homozygous B-hLTA/hTNFA/hLTB mice, as the antibody is crossly reactive with human LTB.
