Strain specific analysis of PAC1R mRNA expression in wild-type C57BL/6 mice and B-hPAC1R mice by RT-PCR. Brain RNA was isolated from wild-type C57BL/6 mice (+/+) and homozygous B-hPAC1R mice (H/H), then cDNA libraries were synthesized by reverse transcription, followed by PCR with mouse or human PAC1R primers. Mouse PAC1R mRNA was detectable only in wild-type C57BL/6 mice. Human PAC1R mRNA was detectable only in homozygous B-hPAC1R mice but not in wild-type mice.

Western blot analysis of PAC1R protein expression in homozygous B-hPAC1R mice. Various tissue lysates were collected from wild-type C57BL/6 mice (+/+) and homozygous B-hPAC1R mice (H/H), and then analyzed by western blot with cross reactive anti-PAC1R antibody. 30 μg (A) /40 μg (B) total proteins were loaded for western blotting analysis. PAC1R was detected in heart, hypothalamus, hippocampus, frontal lobe, cerebral cortex, cerebellum and brain of homozygous B-hPAC1R mice and wild-type mice.