Basic Information
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Gene targeting strategy
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Gene targeting strategy for B-hHCAR1 mice.
The exon 1 of mouse Hcar1 gene that encodes the full-length protein was replaced by human HCAR1 exon 1 in B-hHCAR1 mice.
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mRNA expression analysis
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Strain specific analysis of HCAR1 gene expression in wild-type C57BL/6 mice and B-hHCAR1 mice by RT-PCR. Mouse Hcar1 mRNA was detectable only in BAT (brown adipose tissue) of wild-type C57BL/6 mice (+/+). Human HCAR1 mRNA was detectable only in homozygous B-hHCAR1 mice (H/H), but not in wild-type mice.
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Protein expression analysis
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Strain specific HCAR1 expression analysis in homozygous B-hHCAR1 mice by flow cytometry. BAT (brown adipose tissue) was collected from wild-type C57BL/6 mice (+/+) and homozygous B-hHCAR1 mice (H/H), and analyzed by western blot with anti-HCAR1 antibody. HCAR1 was both detectable in wild-type mice and homozygous B-hHCAR1 mice (H/H) due to the cross-reactivity of antibodies.
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Summary
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mRNA expression analysis:
Mouse Hcar1 mRNA was detectable in BAT of wild-type (+/+). Human HCAR1 mRNA was detectable in B-hHCAR1 mice (H/H) mice but not in wild-type mice.
Protein expression analysis:
HCAR1 was both detectable in wild-type mice and homozygous B-hHCAR1 mice due to the cross-reactivity of antibodies.
