Strain specific FZD5 expression analysis in homozygous B-hFZD5 mice. Various tissue lysates were collected from wild-type C57BL/6 mice(+/+) and homozygous B-hFZD5 mice(H/H), and then analyzed by western blot with anti-FZD5 antibody. 40 μg total protein was loaded for western blotting analysis. FZD5 was detected in brain, as the antibody recognizes both human and mouse FZD5.
Strain specific analysis of FZD5 mRNA expression in wild-type C57BL/6 mice and B-hFZD5 mice by RT-PCR. Brain RNA were isolated from wild-type C57BL/6 mice (+/+) and homozygous B-hFZD5 mice (H/H), then cDNA libraries were synthesized by reverse transcription, followed by PCR with mouse or human FZD5 primers. Mouse Fzd5 mRNA was detectable only in wild-type C57BL/6 mice. Human FZD5 mRNA was detectable only in homozygous B-hFZD5 mice but not in wild-type mice. And the PCR products were confirmed with sanger sequencing.