Figure 1: PLCδ1^−/− mice show granulocytosis in non-haematopoietic cell intrinsic manner.

(a) Representative FACS profiles of CD11b⁺ Gr-1⁺ granulocytes in peripheral blood leukocytes (PBL), spleen, and bone marrow (BM) (n=14). Granulocytes in bone marrow were categorized into immature (CD11b⁺ Gr-1^low) and mature (CD11b⁺ Gr-1^high) subsets. (b) Macroscopic appearance of ILNs. Scale bar, 2 mm. (c) Representative FACS profiles of B220⁺ and Ter119⁺ cells in the BM (n=7–12). (d) BrdU incorporation in immature granulocytes (CD11b⁺ Gr-1^low) in the BM. Data are expressed as the relative mean fluorescence intensity (MFI)±s.e.m. (MFI of PLCδ1^+/− mice=1) (n=5). (e) Populations of myeloid progenitor-rich Lin⁻ Sca-1⁻ c-Kit⁺ cells in the BM. Mean±s.e.m. (n=8). (f) In vitro colony-forming assay. The numbers of colony-forming cells per 6×10⁴ cells at 12 days after plating are displayed. Mean±s.e.m. (n=4). (g) Haematopoietic chimerism analysis. Chimerism of the PBL, spleen, and BM were determined in transplanted mice by FACS analysis of CD45.1⁻ cells. Mean percentage±s.e.m. Six recipients in each group. Three donor mice per genotype were used. (h) Representative FACS profiles of CD45.1⁻ CD11b⁺ Gr-1⁺ granulocytes in the PBL, spleen, and BM of CD45.1⁺ congenic recipients reconstituted with PLCδ1^+/− or PLCδ1^−/− BM at 4 weeks post-transplantation. (n=6). Plots shown are gated on CD45.1⁻ cells. (i) Macroscopic appearance of the ILNs. Scale bar, 2 mm. Mice used in all experiments were 8–12 weeks old. Statistical significance was assessed using a Student′s t-test. *P<0.05; **P<0.01.