德晋贵宾厅

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.

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-recognized in both human and mouse CSPG5. It is impossible to conclude from the data that the observed three bands are CSPG5 three isoforms.

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.