Basic Information
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Targeting strategy
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Gene targeting strategy for B-hIL17RB/hIL25 mice.
The exons 2-10 of mouse Il17rb gene that encode the extracellular domain were replaced by human IL17RB exons 2-10 in B-hIL17RB/hIL25 mice. The exons 2-3 of mouse Il25 gene that encode the full-length protein were replaced by human IL25 exons 2-7 in B-hIL17RB/hIL25 mice.
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mRNA expression analysis
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Strain specific analysis of IL17RB and IL25 gene expression in wild type (WT) mice and B-hIL17RB/hIL25 mice by RT-PCR.
Mouse Il25 mRNA was detectable only in ovary of WT mice (+/+). Human IL25 mRNA was detectable only in homozygous B-hIL17RB/hIL25 mice (H/H;H/H) but not in WT mice (+/+). Mouse Il17rb mRNA was detectable only in thymus of WT mice (+/+). Human IL17RB mRNA was detectable only in homozygous B-hIL17RB/hIL25 mice (H/H;H/H) but not in WT mice (+/+).
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Protein expression analysis in macrophages
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Strain specific IL17RB expression analysis in homozygous B-hIL17RB/hIL25 mice by flow cytometry.
Splenocytes were collected from wild type (WT) mice (+/+) and homozygous B-hIL17RB/hIL25 mice (H/H;H/H), and analyzed by flow cytometry with anti-IL17RB antibody. IL17RB was detectable in WT mice (+/+) and homozygous B-hIL17RB/hIL25 mice (H/H;H/H) due to the cross-reactivity of antibodies.
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Function assay
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Splenocytes from wild type (WT) mice and homozygous B-hIL17RB/hIL25 mice produce IL-5 in response to IL-25 stimulation.
Splenocytes were collected from WT mice (+/+) and homozygous B-hIL17RB/hIL25 mice (H/H) and stimulated with mouse IL-25 or human IL-25. IL-5 concentrations were measured in cell culture supernatants by ELISA. Bars represent the mean ±SEM from three mice. The results showed that cultured splenocytes from WT mice and homozygous B-hIL17RB/hIL25 mice could produce IL-5 in response to human IL-25 or mouse IL-25 stimulation.
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Summary
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mRNA expression analysis:
Mouse Il25 mRNA was detectable only in ovary of WT mice (+/+). Human IL25 mRNA was detectable only in homozygous B-hIL17RB/hIL25 mice (H/H;H/H) but not in WT mice (+/+). Mouse Il17rb mRNA was detectable only in thymus of WT mice (+/+). Human IL17RB mRNA was detectable only in homozygous B-hIL17RB/hIL25 mice (H/H;H/H) but not in WT mice (+/+).
Protein expression analysis:
IL17RB was detectable on macrophages of WT mice (+/+) and homozygous B-hIL17RB/hIL25 mice (H/H;H/H) due to the cross-reactivity of antibodies.
Function assay:
Cultured splenocytes from WT mice and homozygous B-hIL17RB/hIL25 mice could produce IL-5 in response to human IL-25 or mouse IL-25 stimulation.
