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Fig. 4 | BMC Biology

Fig. 4

From: Phosphatidic acid-dependent localization and basal de-phosphorylation of RA-GEFs regulate lymphocyte trafficking

Fig. 4

Impaired retention of RA-GEF-deficient B cells within the bone marrow. a (Top) Representative B220 and IgM profiles of B220+-gated bone marrow cells from WT or RA-GEF B DKO (DKO) mice. (Middle) Cell numbers of IgMlow or IgMhigh, B220low immature B cells, IgMlow, B220high mature B cells in the bone marrow (n = 5). *1P < 0.001, *2P < 0.001, *3P < 0.001 versus corresponding WT cells. (Bottom) Representative CXCR4 and VLA-4 profiles of B220+ bone marrow cells. b (Top) Representative B220 and IgM profiles of B220+-gated blood cells from WT or DKO mice. (Middle) Representative IgM and IgD profiles of B220+-gated blood cells. (Bottom) Cell numbers of IgMhigh, IgDlow immature and IgMlow, IgD high mature B cells from B220+-gated blood cells (n = 5). *1P < 0.001, *2P < 0.001 versus corresponding WT cells. c (Top) Representative IgM and CD43 profiles of B220low, IgM-gated cells (progenitor B cells) from the bone marrow of WT or DKO mice. (Bottom) Cell numbers of IgM, CD43low pre-B and IgM, CD43 high pro-B cells from B220low, IgM-gated bone marrow cells (n = 5). *P < 0.001 versus corresponding WT cells. d (Top) Representative CD21 and CD23 profiles of B220+-gated spleen cells from WT or DKO mice. (Bottom) Cell numbers of CD21, CD23 immature, CD21high, CD23 low marginal zone (MZB) and CD21high, CD23 high follicular (FOB) B cells from B220+-gated spleen cells (n = 3). *1P < 0.001, *2P < 0.001 versus corresponding WT cells. e (Left) Lysates from WT or DKO B cells stimulated with CXCL12 at the indicated times were subjected to the pull-down assay. Bound Rap1 (Rap1-GTP) and total Rap1 were detected with anti-Rap1. (Right upper) Displacement of WT or DKO B cells was measured on VCAM-1 with or without CXCL12 (n = 30). (Lower) Representative tracks of WT or DKO B cells with CXCL12 are shown. Each line represents a single-cell track. Each bar graph represents the means ± SEM

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