Targeting strategy
Gene targeting strategy for B-hIL10RA/hIL10RB mice. The exons 1-6 of mouse Il10ra gene that encode the extracellular protein were replaced by human IL10RA exons 1-6 in B-hIL10RA/hIL10RB mice. The exons 2-6 of mouse Il10rb gene that encode the extracellular protein were replaced by human IL10RB exons 2-6 in B-hIL10RA/hIL10RB mice.
mRNA expression analysis
Strain specific analysis of IL10RB mRNA expression in wild-type C57BL/6 mice and B-hIL10RA/hIL10RB mice by RT-PCR. Mouse Il10rb mRNA was detectable only in splenocytes of wild-type C57BL/6 mice (+/+). Human IL10RB mRNA was detectable only in homozygous B-hIL10RA/hIL10RB mice (H/H, H/H), but not in wild-type mice.
Protein expression analysis
Strain specific IL10RA and IL10RB expression analysis in homozygous B-hIL10RA/hIL10RB mice by flow cytometry. Dendritic cells from spleen were collected from wild-type C57BL/6 mice (+/+) and homozygous B-hIL10RA/hIL10RB mice (H/H, H/H), and analyzed by flow cytometry with species-specific anti-IL10RA and anti-IL10RB antibody. Mouse IL10RA was detectable in wild-type mice. Human IL10RA and human IL10RB were exclusively detectable in homozygous B-hIL10RA/hIL10RB mice but not in wild-type mice.
Strain specific IL10RA and IL10RB expression analysis in homozygous B-hIL10RA/hIL10RB mice by flow cytometry. Macrophages from spleen were collected from wild-type C57BL/6 mice (+/+) and homozygous B-hIL10RA/hIL10RB mice (H/H, H/H), and analyzed by flow cytometry with species-specific anti-IL10RA and anti-IL10RB antibody. Mouse IL10RA was detectable in wild-type mice. Human IL10RA and human IL10RB were exclusively detectable in homozygous B-hIL10RA/hIL10RB mice but not in wild-type mice.
Strain specific IL10RA and IL10RB expression analysis in homozygous B-hIL10RA/hIL10RB mice by flow cytometry. Peritoneal exudative macrophages were collected from wild-type C57BL/6 mice (+/+) and homozygous B-hIL10RA/hIL10RB mice (H/H, H/H), and analyzed by flow cytometry with species-specific anti-IL10RA and anti-IL10RB antibody. Mouse IL10RA was detectable in wild-type mice. Human IL10RA and human IL10RB were exclusively detectable in homozygous B-hIL10RA/hIL10RB mice but not in wild-type mice.
