Figure 5.

Disturbed hematopoiesis in the bone marrow of MyD88-deficent mice. Bone marrow cells obtained from femurs of wild type (wt), Btk-ko, MyD88-ko and Btk/MyD88-deficient mice (DKO) were counted with a ViCell XR Cell Viability Analyzer and analyzed by flow cytometry to distinguish (A) LT-HSC (lin^- Sca1⁺ Kit⁺ CD48^- CD150⁺), (B) multipotent progenitors (lin^- Sca1⁺ Kit⁺ CD48^- CD150^-), (C) common lymphoid progenitors (lin^- Sca1^med Kit^med IL-7Ra⁺), (D) common myeloid progenitors (lin^- Sca1^- Kit⁺ CD34⁺ CD16^med), (E) granulocyte/macrophage progenitor (lin^- Sca1^- Kit⁺ CD34⁺ CD16^high) and (F) megakaryocyte/erythrocyte progenitor (lin^- Sca1^- Kit⁺ CD34^- CD16^-). Data presented are mean values (±SD) (n=6). To analyze myeloid differentiation potential of granulocyte-macrophage progenitors in vitro, 500 GMP were sorted out of individual mice from each genotype and seeded in methylcellulose media supplemented with SCF, IL3 and GM-CSF. G: After 6 days in culture colony-forming units (CFU) were counted. Data presented are the mean values (±SD) (n=5). H: The cell number per CFU was calculated. I: Twenty to thirty individual CFU per biological replicate were processed for cytospins plus Pappenheim staining and analyzed for the cell content by morphology. CFU-M=more than 90% of the cells were macrophages, CFU-G=more than 90% of cells were granulocytes, CFU-GM=the content of macrophages and granulocytes was in between 20 to 80%. The data are presented as mean values (±SEM) (n=5). *P ≤ 0.05, **P ≤ 0.005 . n represents the number of biological replicates.