Basic Information
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Targeting strategy
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Gene targeting strategy for B-Cd11b-EGFP-DTR-Luc mice.
CD11b co-express EGFP, the DTR for depletion of CD11b+ cells, and the luciferase for in vivo bioluminescent imaging (BLI) of CD11b+ cells. EGFP-DTR-Luc expression is evident in CD11b+ cells.
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Protein expression analysis
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Strain specific EGFP expression analysis in heterozygous B-Cd11b-EGFP-DTR-Luc mice by flow cytometry. Splenocytes were collected from wild-type C57BL/6 mice (+/+) and heterozygous B-Cd11b-EGFP-DTR-Luc mice (Mut/+), and analyzed by flow cytometry. EGFP was not detectable in B cells, T cells, while EGFP was detectable in NK cells and CD11b+ cells of heterozygous mice.
Strain specific EGFP expression analysis in heterozygous B-Cd11b-EGFP-DTR-Luc mice by flow cytometry. Splenocytes were collected from wild-type C57BL/6 mice (+/+) and heterozygous B-Cd11b-EGFP-DTR-Luc mice (Mut/+), and analyzed by flow cytometry. EGFP was detectable in DCs, neutrophils, monocytes and macrophages of heterozygous mice.
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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-Cd11b-EGFP-DTR-Luc mice (Mut/+) (n=3, 9-week-old) injected with PBS or DT (50 ng/g body weight) for three 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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CD11b+ 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-Cd11b-EGFP-DTR-Luc mice (Mut/+) (n=3, 9-week-old) injected with PBS or DT (50 ng/g body weight) for three consecutive days. Flow cytometry analysis of the splenocytes was performed to assess the frequencies of CD11b+ EGFP+ cells and EGFP+ cells in total CD45+ cells. The frequencies of CD11b+ EGFP+ cells (A) and EGFP+ cells (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-Cd11b-EGFP-DTR-Luc mice (Mut/+) (n=3, 9-week-old) injected with PBS or DT (50 ng/g body weight) for three consecutive days. Flow cytometry analysis of the splenocytes was performed to assess the frequencies of B cells, T cells, NK cells and DCs in total CD45+ cells. The frequencies of NK cells and DCs (B) were decreased in heterozygous mice injected with DT, while the frequencies of B cells and T cells (A) were not changed.
Frequencies of neutrophils, monocytes and macrophages in spleen by flow cytometry. Splenocytes were isolated from wild-type C57BL/6 mice (+/+) and heterozygous B-Cd11b-EGFP-DTR-Luc mice (Mut/+) (n=3, 9-week-old) injected with PBS or DT (50 ng/g body weight) for three consecutive days. Flow cytometry analysis of the splenocytes was performed to assess the frequencies of neutrophils, monocytes and macrophages in total CD45+ cells. The frequencies of neutrophils (A), monocytes (B) and macrophages (B) were decreased in heterozygous mice injected with DT.
