Fig 1: Immune response and migration of B cells.A. Littermate WT control and RhoF KO mice were immunized with 20 μg of TNP-LPS (200 μl/mouse; WT, 6–8 weeks old, n = 7, male: 86%, KO, 6–9 weeks old, n = 5, male: 80%) or PBS (200 μl/mouse; WT, 6–15 weeks old, n = 9, male: 78%, KO, 6–18 weeks old, n = 9, male: 78%) by injection into the peritoneal cavity and bled at designated time points. Antibody titers of TNP-specific IgM were measured by ELISA. Circles: WT. Crosses: RhoF KO.B. Antibody titers of TNP-specific IgG3 at the indicated time point were measured by ELISA. The samples were same as those for TNP-specific IgM in Figure 5A. Circles: WT. Crosses: RhoF KO.C. Migration assay of MZ B cells. Total splenocytes (106 cells/well) were placed in the upper chamber of Transwell, which was set on the lower chamber with 10% FBS/RPMI 1640 medium with or without BLC (800 ng/ml, BLC) or SDF-1α (200 ng/ml, SDF1). In this experiment, FACS analysis (with anti-CD21, IgM, and CD23) was performed using the input cells and cells in the lower chamber after incubation, to calculate the percentage of MZ B cells that migrated into the lower chamber. The data are shown as mean ± standard deviation (11–12 weeks old, n = 4, male: 50%) from two independent experiments. White bars: WT. Black bars: RhoF KO.