Figure 5.

CNB1-deficient natural killer- natural killer T- and γδ T-cell development in a competitive setting. (A) CD45.2 chimaerism in NK cells in the spleen and in NK T cells in the thymus normalized to myeloid reconstitution. Each symbol represents an individual mouse and the bars represents the average; P refers to two-tailed t-test. (B) Flow cytometry analysis of γδ T cell in the thymus and spleen of a mouse reconstituted with CD45.2⁺ control or CNB1-deficient LSK progenitors and helper CD45.1⁺ bone marrow. Before analysis, γδ T cells were enriched with anti-PE magnetic beads. The percentage of γδ T cells after gating on CD45.1⁺- and CD45.2⁺-positive thymocytes is shown. (C) CD45.2 chimaerism in γδ T cells in the thymus and spleen normalized to myeloid compartment. Each symbol represents an individual mouse and the bars represents the average; P refers to two-tailed t-test. (D) Absolute numbers of αβ T cells in the spleen in mice of the indicated phenotype (n=6, mean±s.d.). (E) Peripheral αβ T cells from mice of the indicated genotype were stimulated in vitro with anti-CD3 and anti-CD28 in the presence or absence of cyclosporine A, and then analysed by flow cytometry for the upregulation of CD40L and BrdU incorporation. (F) Summary of calcineurin-dependent and -independent haematopoietic lineages. BrdU, 5-bromodeoxyuridine; CLP, common lymphoid progenitor; CMP, common myeloid progenitor; GMP, granulocyte-macrophage progenitor; HSC, haematopoietic stem cell; LSK, Lineage^−/lowSca1^highcKit^high; MEP, megakaryocyte progenitor (adapted from Iwasaki & Akashi, 2007); NK, natural killer; PE, phycoerythrin; TCR, T-cell receptor.