American Society of Hematology

Fig. 6.

$Fig. 6. Analysis of primitive and definitive hematopoietic potential ofRunx1+/+, Runx1+/−, and Runx1−/− embryos and ES cells. / (A) Primitive erythroid, definitive erythroid, and macrophage colonies generated by yolk sac cells fromRunx1+/+ (+/+), Runx1+/−(+/−), and Runx1−/− (−/−) embryos. The stage of development (number of somite pairs) is indicated. The number of embryos analyzed at each stage is as follows: 4-6sp: 3 +/+, 2 +/−, 2−/−; 7-9sp: 3 +/+, 3 +/−, 4−/−; 10-12sp: 3 +/+, 12 +/−, 3−/−; and 13-15sp: 1 +/+, 4 +/−, 2−/−. (B) Primitive erythroid precursors generated by Runx1+/+ (+/+),Runx1+/− (+/−), andRunx1−/− (−/−) EBs. Numbers are days of EB differentiation.−/−A and −/−B represent 2 independent clones ofRunx1−/− ES cells. Bars, where visible, represent standard error of the mean number of colonies from at least 3 cultures. (C) Definitive hematopoietic potential ofRunx1+/+ (+/+),Runx1+/− (+/−), andRunx1−/− (−/−) EBs at day 6 of differentiation. Macrophage (MAC), definitive erythroid (ERY/D), and multilineage (MIX) colonies were scored. Bars represent standard error of the mean number of colonies from at least 3 cultures. (D) FACS profile of day 4.75 Runx1+/Z EB cells stained with FDG (left panel). Negative (Neg) and positive (Pos) fractions were isolated and assayed for precursor potential. Number of primitive erythroid (ERY/P) and macrophage (MAC) colonies generated by the sorted fractions is shown in right panel. Bars represent standard error of the mean number of colonies from at least 3 cultures. (E) X-gal staining of Runx1+/+ (+/+) andRunx1+/Z (+/Z) primitive erythroid (ERY/P) and macrophage (MAC) colonies following 4 days of culture.$

Analysis of primitive and definitive hematopoietic potential ofRunx1^+/+, Runx1^+/−, and Runx1^−/− embryos and ES cells.

(A) Primitive erythroid, definitive erythroid, and macrophage colonies generated by yolk sac cells fromRunx1^+/+ (+/+), Runx1^+/−(+/−), and Runx1^−/− (−/−) embryos. The stage of development (number of somite pairs) is indicated. The number of embryos analyzed at each stage is as follows: 4-6sp: 3 +/+, 2 +/−, 2−/−; 7-9sp: 3 +/+, 3 +/−, 4−/−; 10-12sp: 3 +/+, 12 +/−, 3−/−; and 13-15sp: 1 +/+, 4 +/−, 2−/−. (B) Primitive erythroid precursors generated by Runx1^+/+ (+/+),Runx1^+/− (+/−), andRunx1^−/− (−/−) EBs. Numbers are days of EB differentiation.−/−A and −/−B represent 2 independent clones ofRunx1^−/− ES cells. Bars, where visible, represent standard error of the mean number of colonies from at least 3 cultures. (C) Definitive hematopoietic potential ofRunx1^+/+ (+/+),Runx1^+/− (+/−), andRunx1^−/− (−/−) EBs at day 6 of differentiation. Macrophage (MAC), definitive erythroid (ERY/D), and multilineage (MIX) colonies were scored. Bars represent standard error of the mean number of colonies from at least 3 cultures. (D) FACS profile of day 4.75 Runx1^+/Z EB cells stained with FDG (left panel). Negative (Neg) and positive (Pos) fractions were isolated and assayed for precursor potential. Number of primitive erythroid (ERY/P) and macrophage (MAC) colonies generated by the sorted fractions is shown in right panel. Bars represent standard error of the mean number of colonies from at least 3 cultures. (E) X-gal staining of Runx1^+/+ (+/+) andRunx1^+/Z (+/Z) primitive erythroid (ERY/P) and macrophage (MAC) colonies following 4 days of culture.

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