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Figure 4


Fig. 4. Impaired definitive hematopoiesis in c-Myc-deficient embryos. (A) Expression of the pan-hematopoietic marker CD45 in control and c-Myc-deficient dorsal aorta (top panels) and fetal liver (bottom panels) at E11.0. (B) FACS analysis showing CD45 expression in E11.0 embryos. Numbers in the CD45+ gates represent percentage of cells per embryo. (C) Quantitative analysis of total CD45+ cells per E11.0 embryos (as defined in B) (**P≤0.001). (D) FACS analysis showing the expression of the stem cell markers Kit (CD117) and AA4.1 (CD93) in CD45+ hematopoietic cells of control (left) and c-Myc-deficient (right) embryos at E11.0. Numbers represent percentages in the CD45+ population. (E) Quantitative analysis showing the percentage of Kit+AA4.1+ (stem/progenitor) cells within the CD45+ population in E11.0 control and Sox2Cre;c-mycflox/flox embryos (*P≤0.01). (F) Quantitative analysis showing the number of CD45+Kit+AA4.1+ cells per E11.0 control and Sox2Cre;c-mycflox/flox embryos. (G) CFU-assay at E10.5. Examples of blast-forming unit-erythroid (BFU-E) and colony-forming unit-macrophage/granulocyte (CFU-M/G) are shown for control embryos. Cells derived from Sox2Cre;c-mycflox/flox embryos failed to give rise to typical colonies, but only produced very rare colonies composed of very small cells (bottom right). (H) Quantitative analysis of total colony number per embryo after 8 days in culture.