Basic Information
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Description
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- B-Lyz2-EGFP-DTR-luciferase mice have the mouse Lyz2 promoter to determine the expression of DTR (Diphtheria toxin receptor), EGFP (enhanced green fluorescent protein) and luciferase.
- Neutrophils, monocytes and macrophages will be traced by EGFP or Luciferase reporting system in B-Lyz2-EGFP-DTR-luciferase mice.
- Neutrophils, monocytes and macrophages will be depleted in B-Lyz2-EGFP-DTR-luciferase mice injected with DT (diphtheria toxin).
- The mice can be used to study the specific role of neutrophils, monocytes and macrophages in immune response.
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Targeting strategy
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Gene targeting strategy for B-Lyz2 EGFP-DTR-luciferase mice.
To avoid disrupting the expression of Lyz2 gene, EGFP-DTR-luciferase will be introduced between the coding sequence of exon4 and UTR.
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Protein expression analysis
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Strain specific EGFP expression analysis in homozygous B-Lyz2-EGFP-DTR-luciferase mice by flow cytometry. Splenocytes were collected from wild-type C57BL/6 mice (+/+) and heterozygous B-Lyz2-EGFP-DTR-luciferase mice (Mut/+). EGFP was detected in CD11b+ cells, macrophages, neutrophils and monocytes of heterozygous mice, but not in wild-type mice.
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LYZ2+ cells depletion analysis
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Frequencies of CD11b+ EGFP+ cells and EGFP+ cells in spleen by flow cytometry. Splenocytes were isolated from wild-type C57BL/6 mice (+/+) and heterozygous B-Lyz2-EGFP-DTR-luciferase mice (Mut/+) (Male, n=3, 8-week-old) injected with PBS or DT (50 ng/g body weight) for two consecutive days. Flow cytometry analysis of the splenocytes was performed to assess the frequencies of CD11b+ EGFP+ cells (A) and EGFP+ cells (B) in total CD45+ cells. The frequencies of EGFP+ cells (A) and CD11b+ EGFP+ cells (B) were decreased in heterozygous mice injected with DT.
Frequencies of neutrophils, monocytes and macrophages in spleen by flow cytometry. Splenocytes were isolated from wild-type C57BL/6 mice (+/+) and heterozygous B-Lyz2-EGFP-DTR-luciferase mice (Mut/+) (Male, n=3, 8-week-old) injected with PBS or DT (50 ng/g body weight) for two consecutive days. Flow cytometry analysis of the splenocytes was performed to assess the frequencies of neutrophils (A), monocytes (B) and macrophages (B) in total CD45+ cells. The frequencies of neutrophils (A), monocytes (B) and macrophages (B) were decreased in heterozygous mice injected with DT.
Frequencies of B cells, T cells, NK cells and DCs in spleen by flow cytometry. Splenocytes were isolated from wild-type C57BL/6 mice (+/+) and heterozygous B-Lyz2-EGFP-DTR-luciferase mice (Mut/+) (Male, n=3, 8-week-old) injected with PBS or DT (50 ng/g body weight) for two consecutive days. Flow cytometry analysis of the splenocytes was performed to assess the frequencies of B cells, T cells (A) and NK cells, DCs (B) in total CD45+ cells. The frequencies of B cells, T cells (A) and NK cells, DCs (B) were not changed in heterozygous mice injected with DT.
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Frequency of leukocyte subpopulations in spleen
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Frequency of leukocyte subpopulations in spleen by flow cytometry. Splenocytes were isolated from wild-type C57BL/6 mice (+/+) and heterozygous B-Lyz2-EGFP-DTR-luciferase mice (Mut/+) (Male, n=3, 8-week-old) injected with PBS or DT (50 ng/g body weight) for two consecutive days. Flow cytometry analysis of the splenocytes was performed to assess the frequency of leukocyte subpopulations. Values are expressed as mean ± SEM.
