Frequency of leukocyte subpopulations in spleen by flow cytometry. Splenocytes were isolated from female wild-type BALB/c mice (n=3, 7-week-old) and homozygous B-Rag2 KO mice(C) (n=3, 7-week-old). A. Flow cytometry analysis of the splenocytes was performed to assess the frequency of leukocyte subpopulations. B. Frequency of T cell subpopulations. The frequency of total T cells, CD4+ T cells, CD8+ T cells, Tregs and B cells were decreased in B-Rag2 KO mice(C). The frequency of NK cells, DCs, neutrophils and macrophages were increased in B-Rag2 KO mice(C) compared to BALB/c mice, demonstrating that Rag2 was successfully knocked out in spleen. Values are expressed as mean ± SEM. Significance was determined by two-way ANOVA test. *P < 0.05, **P < 0.01, ***p < 0.001.

Frequency of leukocyte subpopulations in blood by flow cytometry. Blood cells were isolated from female wild-type BALB/c mice (n=3, 7-week-old) and homozygous B-Rag2 KO mice(C) (n=3, 7-week-old). A. Flow cytometry analysis of the blood cells was performed to assess the frequency of leukocyte subpopulations. B. Frequency of T cell subpopulations. The frequency of total T cells, CD4+ T cells, CD8+ T cells, Tregs and B cells were decreased in B-Rag2 KO mice(C). The frequency of NK cells, DCs, neutrophils and macrophages were increased in B-Rag2 KO mice(C) compared to BALB/c mice, demonstrating that Rag2 was successfully knocked out in blood. Values are expressed as mean ± SEM. Significance was determined by two-way ANOVA test. *P < 0.05, **P < 0.01, ***p < 0.001.

Frequency of leukocyte subpopulations in bone marrow by flow cytometry. Bone marrow cells were isolated from female wild-type BALB/c mice (n=3, 7-week-old) and homozygous B-Rag2 KO mice(C) (n=3, 7-week-old). A. Flow cytometry analysis of the bone marrow cells was performed to assess the frequency of leukocyte subpopulations. B. Frequency of T cell subpopulations. The frequency of total T cells, CD4+ T cells, CD8+ T cells, Tregs and B cells were decreased in B-Rag2 KO mice(C). The frequency of NK cells, DCs, neutrophils and macrophages were increased in B-Rag2 KO mice(C) compared to BALB/c mice, demonstrating that Rag2 was successfully knocked out in bone marrow. Values are expressed as mean ± SEM. Significance was determined by two-way ANOVA test. *P < 0.05, **P < 0.01, ***p < 0.001.

The size and weight of thymus and spleen of B-Rag2 KO mice(C) were significantly reduced compared to that of wild-type BALB/c mice. Thymuses, spleens and lymph nodes were isolated from wild-type BALB/c mice and B-Rag2 KO mice(C) (n=3, 7-week-old). A. Gross anatomy of thymuses; B. The size of spleen, thymus and lymph node; C. Comparison of the weight of spleen and thymus in wild-type BALB/c mice and B-Rag2 KO mice(C). Results showed that the size and weight of spleens and thymuses in B-Rag2 KO mice(C) were significantly reduced compared to that in wild-type BALB/c mice. Lymph nodes were not visualized in B-Rag2 KO mice(C).