德晋贵宾厅

Gene targeting strategy for B-hCDH3 mice.
The exons 4-13 of mouse Cdh3 gene that encode the extracellular domain were replaced by human CDH3 exons 4-13 in B-hCDH3 mice.

Simple Western analysis of CDH3 protein expression in homozygous B-hCDH3 mice. Various tissue lysates were collected from wild-type C57BL/6 (+/+) mice and homozygous B-hCDH3 mice (H/H), and then analyzed by Simple Western with anti-CDH3 antibody. 1mg/mL total proteins were loaded for simple western analysis. CDH3 protein was detected in the lung, spleen,thymus gland and skin of wild-type C57BL/6 mice and homozygous B-hCDH3 mice,as the antibody is cross-recognize both human and mouse CDH3. Human CDH3 was detected in the lung, spleen,thymus gland and skin of homozygous B-hCDH3 mice.

Strain specific analysis of CDH3 mRNA expression in wild-type C57BL/6 mice and B-hCDH3 mice by RT-PCR. Thymus gland RNA were isolated from wild-type C57BL/6 mice (+/+) and homozygous B-hCDH3 mice (H/H), then cDNA libraries were synthesized by reverse transcription, followed by PCR with mouse or human CDH3 primers. Mouse Cdh3 mRNA was detectable only in wild-type C57BL/6 mice. Human CDH3 mRNA was detectable only in homozygous B-hCDH3 mice but not in wild-type mice.

Western blot analysis of CDH3 protein expression in homozygous B-hCDH3 mice. Oesophagus and tongue tissue lysates were collected from wild-type C57BL/6N mice (+/+) and homozygous B-hCDH3 mice (H/H), and then analyzed by western blot with species-specific anti-CDH3 antibody(anti-mouse CDH3,R&D,AF761-SP; anti-human CDH3,R&D,MAB861-SP). 40 μg total proteins were loaded for western blotting analysis. Mouse CDH3 was only detectable in the oesophagus and tongue of wild-type C57BL/6 mice .Human CDH3 was exclusively detectable in the oesophagus and tongue of homozygous B-hCDH3 mice.