Targeting strategy
Gene targeting strategy for B-hEREG mice. The exons 2~4 of mouse Ereg gene were replaced by human EREG exons 2~4 in B-hEREG mice.
mRNA expression analysis
Strain specific analysis of EREG mRNA expression in wild-type C57BL/6 mice and B-hEREG mice by RT-PCR. Small intestine RNA were isolated from wild-type C57BL/6 mice (+/+) and homozygous B-hEREG mice (H/H), then cDNA libraries were synthesized by reverse transcription, followed by PCR with mouse or human EREG primers. Mouse Ereg mRNA was detectable only in wild-type C57BL/6 mice. Human EREG mRNA was detectable only in homozygous B-hEREG mice but not in wild-type mice.
Protein expression analysis
Strain specific EREG expression analysis in homozygous B-hEREG mice by ELISA. Colon homogenate were collected from wild-type mice C57BL/6 mice (+/+) and homozygous B-hEREG mice (H/H), and analyzed by ELISA with species-specific EREG ELISA kit. Human EREG was exclusively detectable in homozygous B-hEREG mice but not in wild-type mice (Each mg of total protein contains approximately 150 pg of EREG).
