B-Lyz2 EGFP-DTR-luciferase mice

Basic Information

Strain name
C57BL/6JNifdc-Lyz2tm1(EGFP-DTR-Luc)Bcgen/Bcgen
Common name
B-Lyz2 EGFP-DTR-luciferase mice
Background
C57BL/6JNifdc
Catalog number
112826

Description

  • 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.

Targeting strategy

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.

Protein expression analysis

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.

LYZ2+ cells depletion analysis

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.

 

Frequency of leukocyte subpopulations in spleen

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.

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