RhoA of the Rho Family Small GTPases Is Essential for B Lymphocyte Development
Figure 1
B cell-specific deletion of RhoA impairs splenic B cell development.
(A) Generation of RhoA−/− B cells. Left, the loxP/Cre-mediated gene targeting strategy to generate the RhoA knockout allele (RhoA−) in B cells. Right, Western blot showing RhoA expression in B220+ B cells purified from bone marrow and spleen of CD19Cre/+; RhoA+/+ (control) and CD19Cre/+; RhoAflox/flox (RhoA−/−) mice. (B) Bone marrow cells from control and RhoA−/− mice were stained with antibodies against B220 and IgM and analyzed by flow cytometry (left). The number of B cell subsets was calculated by multiplying the total number of bone marrow cells by the percentage of each subset of cells (right). n = 5. (C) Splenocytes from control and RhoA−/− mice were stained with antibodies against B220, CD21 and CD23 and analyzed by flow cytometry (left). The number of B cell subsets was calculated by multiplying the total number of splenocytes by the percentage of each subset of cells (right). T: transitional B cells, FO B: follicular B cells, and MZ B: marginal zone B cells. n = 5. (D) Spleen sections from control and RhoA−/− mice, stained with hematoxylin and eosin. Data are representative of 3 mice. Error bars represent mean ± SD. **p<0.01. Statistical analysis was performed using a Student's unpaired t-test with a two-tailed distribution.