Basic Information
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Targeting strategy
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Gene targeting strategy for B-hOSM/hOSMR mice.
The exons 1-2 of mouse Osm gene that encode the full-length protein were replaced by human OSM exons 1-3 in B-hOSM/hOSMR mice. The coding sequences of human OSMR extracellular region and mouse Osmr intracellular region were inserted into mouse Osmr gene in B-hOSM/hOSMR mice.
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mRNA expression analysis
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Strain specific analysis of OSMR gene expression in wild type and B-hOSM/hOSMR mice by RT-PCR and RT-qPCR. (A) OSMR mRNA expression. Mouse Osmr mRNA was exclusively detectable in kidney of wild-type mice (+/+). Human OSMR mRNA was only detectable in homozygous B-hOSM/hOSMR mice (H/H), but not in wild type mice. (B) OSMR mRNA expression level. The results show that introduction of hOSMR in place of its mouse counterpart does not change the mRNA expression level.
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Protein expression analysis
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Strain specific OSMR expression analysis in wild-type C57BL/6 mice and B-hOSM/hOSMR mice by western blot. Kidney tissues were collected from wild-type C57BL/6 mice (+/+) and homozygous B-hOSM/hOSMR mice (H/H), and analyzed by western blot with a known cross-reactive anti-OSMR antibody. 293T cells were used as positive control (PC). The OSMR protein can be detected in wild-type C57BL/6 mice and homozygous B-hOSM/hOSMR mice.
Strain specific OSM expression analysis in homozygous B-hOSM/hOSMR mice by ELISA. Spleen were isolated from wild-type mice (+/+) and homozygous B-hOSM/hOSMR mice (H/H) and stimulated with 2ug/mL anti-mouse CD3 and 5ug/mL anti-mouse CD28 in vitro, cell culture supernates were analyzed by ELISA with species-specific OSM ELISA kit. Mouse OSM was detectable in wild-type mice. Human OSM was exclusively detectable in homozygous B-hOSM/hOSMR mice but not in wild-type mice.
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Summary
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mRNA expression analysis:
Human OSMR mRNA was detectable in B-hOSM/hOSMR mice but not in wild-type mice.
Protein expression analysis:
Human OSM and OSMR were detectable in B-hOSM/hOSMR mice.
