Basic Information
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Targeting strategy
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Gene targeting strategy for B-hLOX1 mice.
The exon 2-6 of mouse LOX-1 gene that encodes the extracellular domain and transmembrane region was replaced by human LOX1 exon 2-6 in B-hLOX1 mice.
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mRNA expression analysis
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Strain specific analysis of LOX1 mRNA expression in wild-type C57BL/6 mice and B-hLOX1 mice by RT-PCR. Lung RNA were isolated from wild-type C57BL/6 mice (+/+) and homozygous B-hLOX1 mice (H/H), then cDNA libraries were synthesized by reverse transcription, followed by PCR with mouse or human LOX1 primers. Mouse Lox1 mRNA was detectable only in wild-type C57BL/6 mice. Human LOX1 mRNA was detectable only in homozygous B-hLOX1 mice but not in wild-type mice. And the PCR products were confirmed with Sanger sequencing.
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Protein expression analysis
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Strain specific LOX1 expression analysis in homozygous B-hLOX1 mice by flow cytometry. Bone marrow-derived dendritic cells(BMDC) were cultured from wild-type C57BL/6 mice (+/+) and homozygous B-hLOX1 mice (H/H) stimulated with LPS (1μg/mL , stimulation for 18 hour), and analyzed by flow cytometry with species-specific anti-LOX1 antibody. Mouse LOX1 was detectable on BMDC of wild-type mice. Human LOX1 was detectable on BMDC of homozygous B-hLOX1 mice.
