Gene targeting strategy for B-hCRTH2 mice. The mouse Ptgdr2 gene that encodes the full length of coding region sequence was replaced by human PTGDR2 counterpart coding sequence in B-hCRTH2 mice.
mRNA and protein expression analysis
Strain specific analysis of CRTH2 gene expression in wild-type mice and B-hCRTH2 mice by RT-PCR. Mouse Crth2 mRNA was detectable only in splenocytes of wild-type mice (+/+). Human CRTH2 mRNA was detectable only in splenocytes of homozygous B-hCRTH2 mice (H/H).
Strain specific CRTH2 expression analysis in homozygous B-hCRTH2 mice by western blot. Spleen and blood tissues were collected from wild-type C57BL/6 mice (+/+) and homozygous B-hCRTH2 mice (H/H), and analyzed by western blot with anti-CRTH2 antibody. CRTH2 was detectable in blood and spleen from wild-type mice and homozygous B-hCRTH2 mice, as the antibody is crossly reactive with human and mouse CRTH2, and shows more nonspecific binding in the spleen.