Basic Information
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mRNA expression analysis
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Species-specific TLR8 and TLR7 gene expression analysis in wild-type and humanized B-hTLR8/hTLR7 mice by RT-PCR. Murine Tlr8 and Tlr7 mRNA was detected in wild-type C57BL/6 (+/+) mice, while human TLR8 and TLR7 mRNA was detected in homozygous B-hTLR8/hTLR7 (H/H) mice.
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Protein expression analysis
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Species-specific TLR7 protein expression analysis in wild-type and humanized B-hTLR8/hTLR7 mice. Spleen-derived dendritic cells were isolated from wild-type C57BL/6 (+/+) and homozygous B-hTLR8/hTLR7 (H/H) mice and analyzed by flow cytometry using species-specific anti-TLR7 antibodies. Murine TLR7 protein was detected in wild-type mice, while human TLR7 protein was detected in B-hTLR8/hTLR7 mice.
Species-specific TLR8 protein expression analysis in wild-type and humanized B-hTLR8/hTLR7 mice. Spleen-derived monocytes were isolated from wild-type C57BL/6 (+/+) and homozygous B-hTLR8/hTLR7 (H/H) mice and analyzed by flow cytometry using species-specific anti-TLR8 antibodies. Murine TLR8 protein was detected in wild-type and B-hTLR8/hTLR7 mice due to the cross-reactive anti-mTLR8 antibody. Human TLR8 protein was only detected in B-hTLR8/hTLR7 mice.
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Immune cell analysis
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Analysis of spleen-derived leukocytes in wild-type and humanized B-hTLR8/hTLR7 mice. Splenocytes were isolated from wild-type C57BL/6 and B-hTLR8/hTLR7 mice (female, n=3, 8-week-old) and analyzed by flow cytometry to assess leukocyte subpopulations. (A) Representative flow cytometry plots. Single live cells were gated on CD45+ cells and used for further analysis as indicated. (B) Percent of T cells, B cells, NK cells, dendritic cells, granulocytes, monocytes and macrophages in B-hTLR8/hTLR7 mice were similar to those in wild-type mice.
Analysis of spleen-derived T cells in wild-type and humanized B-hTLR8/hTLR7 mice. Spleen-derived T cells were isolated from wild-type C57BL/6 and B-hTLR8/hTLR7 mice (female, n=3, 8-week-old) and analyzed by flow cytometry to assess leukocyte subpopulations. (A) Representative flow cytometry plots. Single live CD45+ cells were gated on TCRβ+ T cells and used for further analysis as indicated. (B) Percent of CD8+ T cells , CD4+ T cells and Tregs in B-hTLR8/hTLR7 mice were similar to those in wild-type mice.
