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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.