• 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.

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.

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.

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 in total CD45+ cells. The frequencies of CD11b+ EGFP+ cells were decreased in heterozygous mice injected with DT.