EGFP expression analysis in leukocytes cell subpopulation
Frequencies of EGFP in leukocytes cell subpopulation from spleen, blood and lymph node by flow cytometry. Splenocytes, blood and lymph node were isolated from wild-type C57BL/6 mice (+/+) and heterozygous B-Cd19-EGFP-DTR-luciferase mice (Mut/+) (n=3, 6-7-week-old). Flow cytometry analysis of the splenocytes, blood and lymph node was performed to assess the frequencies of EGFP+ cells in leukocytes cell subpopulation from spleen(A), blood (B) and lymph node(C). EGFP+ cells was expressed at a high level in B cells from spleen, blood and lymph node in heterozygous mice. And there was no leaky expression of EGFP in other cell subpopulation.
CD19+ cells depletion analysis
Frequencies of CD19+; EGFP cells in spleen, blood and lymph node by flow cytometry. Splenocytes, blood and lymph node were isolated from wild-type C57BL/6 mice (+/+) and heterozygous B-Cd19-EGFP-DTR-luciferase mice (Mut/+) (n=3, 6-7-week-old) injected with DT (50 ng per g body weight) for four consecutive days. Flow cytometry analysis of the splenocytes, blood and lymph node was performed to assess the frequencies of CD19+; EGFP+ cells in total CD45+ cells. The signal for CD19+;EGFP+ cells in spleen(A), in blood (B) and lymph node(C) were significantly decreased in heterozygous mice injected with DT.
CD19+ cells depletion analysis in spleen
Frequencies of CD19+ cells and EGFP+ cells in spleen by flow cytometry. Splenocytes were isolated from wild-type C57BL/6 mice (+/+) and heterozygous B-Cd19-EGFP-DTR-luciferase mice (Mut/+) (n=3, 6-7-week-old) injected with DT (50 ng per g body weight) for four consecutive days. Flow cytometry analysis of the splenocytes was performed to assess the frequencies of CD19+ cells and EGFP+ cells in total CD45+ cells. The signal for CD19+ cells(A) and EGFP+ cells(B) were decreased in heterozygous mice injected with DT. C. Percentages of CD19+ cells and EGFP+ cells in total CD45+ populations. The frequencies of CD19+ cells and EGFP+ cells were significantly decreased in heterozygous mice after DT injection.
CD19+ cells depletion analysis in blood
Frequencies of CD19+ cells and EGFP+ cells in blood by flow cytometry. Blood were isolated from wild-type C57BL/6 mice (+/+) and heterozygous B-Cd19-EGFP-DTR-luciferase mice (Mut/+) (n=3, 6-7-week-old) injected with DT (50 ng per g body weight) for four consecutive days. Flow cytometry analysis of the blood was performed to assess the frequencies of CD19+ cells and EGFP+ cells in total CD45+ cells. The signal for CD19+ cells(A) and EGFP+ cells(B) were decreased in heterozygous mice injected with DT. C. Percentages of CD19+ cells and EGFP+ cells in total CD45+ populations. The frequencies of CD19+ cells and EGFP+ cells were significantly decreased in heterozygous mice after DT injection.
CD19+ cells depletion analysis in lymph node
Frequencies of CD19+ cells and EGFP+ cells in lymph node by flow cytometry. Lymph node were isolated from wild-type C57BL/6 mice (+/+) and heterozygous B-Cd19-EGFP-DTR-luciferase mice (Mut/+) (n=3, 6-7-week-old) injected with DT (50 ng per g body weight) for four consecutive days. Flow cytometry analysis of the lymph node was performed to assess the frequencies of CD19+ cells and EGFP+ cells in total CD45+ cells. The signal for CD19+ cells(A) and EGFP+ cells(B) were decreased in heterozygous mice injected with DT. C. Percentages of CD19+ cells and EGFP+ cells in total CD45+ populations. The frequencies of CD19+ cells and EGFP+ cells were significantly decreased in heterozygous mice after DT injection.
B cells depletion analysis
Frequencies of B cells in spleen, blood and lymph node by flow cytometry. Splenocytes, blood and lymph node were isolated from wild-type C57BL/6 mice (+/+) and heterozygous B-Cd19-EGFP-DTR-luciferase mice (Mut/+) (n=3, 6-7-week-old) injected with DT (50 ng per g body weight) for four consecutive days. Flow cytometry analysis of the splenocytes, blood and lymph node was performed to assess the frequencies of B cells in total CD45+ cells. The signal for B cells in spleen(A), blood (B) and lymph node(C) were significantly decreased in heterozygous mice injected with DT.
Frequencies of EGFP in B cells from spleen, blood and lymph node by flow cytometry. Splenocytes, blood and lymph node were isolated from wild-type C57BL/6 mice (+/+) and heterozygous B-Cd19-EGFP-DTR-luciferase mice (Mut/+) (n=3, 6-7-week-old) injected with DT (50 ng per g body weight) for four consecutive days. Flow cytometry analysis of the splenocytes, blood and lymph node was performed to assess the frequencies of EGFP cells in total B cells. The signal for EGFP in B cells in spleen(A), blood (B) and lymph node(C) were significantly decreased in heterozygous mice injected with DT.
Analysis of B cells depletion in spleen, blood and lymph node by flow cytometry. Splenocytes, blood and lymph node were isolated from wild-type C57BL/6 mice (+/+) and heterozygous B-Cd19-EGFP-DTR-luciferase mice (Mut/+) (n=3, 6-7-week-old) injected with DT (50 ng per g body weight) for four consecutive days. The percentages of B cells in total CD45 cells from spleen, blood and lymph node(A) and EGFP+ cells in B cells from spleen, blood and lymph node(B) were significantly decreased 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 B-Cd19-EGFP-DTR-luc mice(n=3, 6-7-week-old). Flow cytometry analysis of the splenocytes was performed to assess the frequency of leukocyte subpopulations. Percentages of T cells, B cells, NK cells, dendritic cells, granulocytes, monocytes and macrophages in heterozygous B-Cd19-EGFP-DTR-luc mice were similar to those in C57BL/6 mice. The frequency of leukocyte subpopulations in blood and lymph node of B-Cd19-EGFP-DTR-luc mice were also comparable to wild-type C57BL/6 mice (Data not shown). Values are expressed as mean ± SEM.
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