Basic Information
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mRNA expression analysis-IL31
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Strain specific analysis of IL31 mRNA expression in wild-type C57BL/6 mice and B-hIL31/hIL31RA/hOSMR/hIL4/hIL4RA mice by RT-PCR. Testis RNA were isolated from wild-type C57BL/6 mice (+/+) and homozygous B-hIL31/hIL31RA/hOSMR/hIL4/hIL4RA mice (H/H), then cDNA libraries were synthesized by reverse transcription, followed by PCR with mouse or human IL31 primers. Mouse IL31 mRNA was only detectable in wild-type mice. Human IL31 mRNA was exclusively detectable in homozygous B-hIL31/hIL31RA/hOSMR/hIL4/hIL4RA mice but not in wild-type mice.
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mRNA expression analysis-IL31RA
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Strain specific analysis of IL31RA mRNA expression in wild-type C57BL/6 mice and B-hIL31/hIL31RA/hOSMR/hIL4/hIL4RA mice by RT-PCR. Testis RNA were isolated from wild-type C57BL/6 mice (+/+) and homozygous B-hIL31/hIL31RA/hOSMR/hIL4/hIL4RA mice (H/H), then cDNA libraries were synthesized by reverse transcription, followed by PCR with mouse or human IL31RA primers. Human IL31RA mRNA was exclusively detectable in homozygous B-hIL31/hIL31RA/hOSMR/hIL4/hIL4RA mice but not in wild-type mice.
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mRNA expression analysis-OSMR
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Strain specific analysis of OSMR mRNA expression in wild-type C57BL/6 mice and B-hIL31/hIL31RA/hOSMR/hIL4/hIL4RA mice by RT-PCR. Kidney RNA were isolated from wild-type C57BL/6 mice (+/+) and homozygous B-hIL31/hIL31RA/hOSMR/hIL4/hIL4RA mice (H/H), then cDNA libraries were synthesized by reverse transcription, followed by PCR with mouse or human OSMR primers. Human OSMR mRNA was exclusively detectable in homozygous B-hIL31/hIL31RA/hOSMR/hIL4/hIL4RA mice but not in wild-type mice.
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Protein expression analysis
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Strain specific IL4 expression analysis in homozygous B-hIL31/hIL31RA/hOSMR/hIL4/hIL4RA mice by ELISA. Serum were collected from wild-type mice C57BL/6 mice (n=3, 6-week-old, female) and homozygous B-hIL31/hIL31RA/hOSMR/hIL4/hIL4RA mice (n=3, 6-week-old, female) that stimulated with anti-mCD3e (7.5 μg/mice, i.p.) and anti-mCD28 (5 μg/mice, i.p.) in vivo for 3 h, and analyzed by ELISA with species-specific IL4 ELISA kit (mIL4 kit: BioLegend 431104; hIL4 kit: BioLegend 430304). Mouse IL4 was only detectable in wild-type mice (A). Human IL4 was exclusively detectable in homozygous mice but not in wild-type mice (B).
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Protein expression analysis in B cells
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Strain specific IL4RA expression analysis in homozygous B-hIL31/hIL31RA/hOSMR/hIL4/hIL4RA mice by flow cytometry. Splenocytes were collected from wild-type C57BL/6 mice and homozygous B-hIL31/hIL31RA/hOSMR/hIL4/hIL4RA mice stimulated with LPS (200 μg/mice, i.p.) for 2 h, and analyzed by flow cytometry with anti-mouse IL4RA antibody (Biolegend, 144803) or anti-human IL4RA antibody (Biolegend, 355005). Mouse IL4RA was only detectable in wild-type mice, while hIL4RA was exclusively detectable in homozygous mice.
