Western blot analysis of GFRA3 protein expression in homozygous B-hGFRA3 mice. Various tissue lysates were collected from wild-type C57BL/6(+/+) mice and homozygous B-hGFRA3 mice (H/H), and then analyzed by western blot with species-specific anti-GFRA3 antibody. 40 μg total proteins were loaded for western blotting analysis. GFRA3 was detected in heart, liver and kidney.

Strain specific analysis of GFRA3 mRNA expression in wild-type C57BL/6 mice and B-hGFRA3 mice by RT-PCR. Heart RNA were isolated from wild-type C57BL/6 mice (+/+) and homozygous B-hGFRA3 mice (H/H), then cDNA libraries were synthesized by reverse transcription, followed by PCR with mouse or human GFRA3 primers. Mouse Gfra3 mRNA was detectable only in wild-type C57BL/6 mice. Human GFRA3 mRNA was detectable only in homozygous B-hGFRA3 mice but not in wild-type mice. And the PCR products were confirmed with Sanger sequencing.