Basic Information
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Targeting strategy
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Gene targeting strategy for B-hCSPG5 mice. The exons 1~3 of mouse Cspg5 gene that encode the extracellular domain were replaced by human CSPG5 exons 1~3 in B-hCSPG5 mice.
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Protein expression analysis
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Western blot analysis of CSPG5 protein expression in homozygous B-hCSPG5 mice. Cortex lysates were collected from C57BL/6 wild-type (+/+) mice and homozygous B-hCSPG5 mice (H/H), and then analyzed by western blot with species-specific anti-CSPG5 antibody. 40 μg total proteins were loaded for western blotting analysis. CSPG5 protein was detected in the cortex of wild-type C57BL/6 mice and homozygous B-hCSPG5 mice, as the antibody is cross-recognize both human and mouse CSPG5. It is impossible to conclude from the data that the observed three bands are CSPG5 three isoforms.
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mRNA expression analysis
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Strain specific analysis of CSPG5 mRNA expression in wild-type C57BL/6 mice and B-hCSPG5 mice by RT-PCR. Cortex RNA was isolated from wild-type C57BL/6 mice (+/+) and homozygous B-hCSPG5 mice(H/H), then cDNA libraries were synthesized by reverse transcription, followed by PCR with mouse or human CSPG5 primers. Mouse Cspg5 mRNA was detectable only in wild-type C57BL/6 mice. Human CSPG5 mRNA was detectable only in homozygous B-hCSPG5 mice but not in wild-type mice.
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Summary
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mRNA expression analysis:
Mouse Cspg5 mRNA was detectable in cortex of wild-type C57BL/6 mice. Human CSPG5 mRNA was detectable in B-hCSPG5 mice.
Protein expression analysis:
CSPG5 protein was detected in the cortex of wild-type C57BL/6 mice and homozygous B-hCSPG5 mice, as the antibody is cross-recognize both human and mouse CSPG5.
