Targeting strategy
Gene targeting strategy for B-hTCIRG1 mice. The exons 2~20 of mouse Tcirg1 gene that encode the full-length protein were replaced by human TCIRG1 exons 2~20 in B-hTCIRG1 mice.
mRNA expression analysis
Strain specific analysis of TCIRG1 mRNA expression in wild-type C57BL/6 mice and B-hTCIRG1 mice by RT-PCR. Spleen RNA were isolated from wild-type C57BL/6 mice (+/+) and homozygous B-hTCIRG1 mice (H/H), then cDNA libraries were synthesized by reverse transcription, followed by PCR with mouse or human TCIRG1 primers. Mouse Tcirg1 mRNA was detectable only in wild-type C57BL/6 mice. Human TCIRG1 mRNA was detectable only in homozygous B-hTCIRG1 mice but not in wild-type mice.
Protein expression analysis
Western blot analysis of TCIRG1 protein expression in homozygous B-hTCIRG1 mice. Various tissue lysates were collected from wild-type C57BL/6 mice (+/+) and homozygous B-hTCIRG1 mice (H/H), and then analyzed by western blot with cross-reactive anti-TCIRG1 antibody. 20 μg total proteins were loaded for western blotting analysis. TCIRG1 was detected in spleen, lymph node, lung, adrenal gland and bone marrow.
