Basic Information
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Targeting strategy
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Gene targeting strategy for B-Cd11c EGFP-DTR-Luc mice.
CD11c co-express EGFP, the DTR for depletion of DCs, and the luciferase for in vivo bioluminescent imaging (BLI) of DCs. EGFP-DTR-Luc expression is evident in CD11c+ cells.
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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-Cd11c-EGFP-DTR-Luc mice (Mut/+) (n=3, 6-8-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. Significance was determined by two-way ANOVA test. *P < 0.05, **P < 0.01, ***p < 0.0001.
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Protein expression analysis
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Frequencies of EGFP+ cells and DCs in spleen by flow cytometry. Splenocytes were isolated from wild-type C57BL/6 mice (+/+) and heterozygous B-Cd11c-EGFP-DTR-Luc mice (Mut/+) (n=3, 6-8-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 EGFP+ cells and DCs. The frequencies of EGFP+ cells and DCs were decreased in heterozygous mice injected with DT.
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Summary
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Protein expression analysis:
- EGFP was only detectable in heterozygous B-Cd11c-EGFP-DTR-Luc mice, but not in wild-type C57BL/6 mice.
- EGFP was only detectable on DCs, NK cells, monocytes and macrophages from spleen of heterozygous B-Cd11c-EGFP-DTR-Luc mice.
Functional analysis:
- The frequencies of DCs and EGFP+ cells were decreased in heterozygous B-Cd11c-EGFP-DTR-Luc mice with injected with DT.
- The frequencies of NK cells and monocytes were decreased in heterozygous B-Cd11c-EGFP-DTR-Luc mice with injected with DT, while the frequencies of neutrophils and macrophages were increased.
