Figure 2
Figure 2. GATA2 expression in normal progenitor-cell fractions. (A) Cord blood CD34+ cells (1 × 105 cells) were cultured for 1 week in medium containing SCF (S), with/without Flt-3L (FL) or Jagged 1/Fc-Protein G-agarose beads (J). Density plot analysis of GATA2 expression and DNA content in the cord blood CD34+ cell fraction. Vertical lines represent borders of 2N population. Bar graphs represent number of total cells (left) and of GATA2+ cells (right) after 1 week's culture. Mean ± standard error of the mean; n = 4. *P < .05 compared with the culture containing SCF. (B) Immunoblot analysis of the cultured cord blood CD34+ fraction. (−) indicates no thymidine addition; T, treated with thymidine for 24 hours; TR, after release from thymidine treatment; and N, treated with nocodazole for 24 hours. (C-G) Lin− fractions of mouse bone marrow cells were cultured with SCF and Jagged 1 (J) or SCF and erythropoietin (E) for 4 days, treated with excess thymidine for additional 24 hours, then released into fresh medium. (C) DNA content histogram. (D) Immunoblot analysis. (E-F) Density plot analysis of GATA1, GATA2, and DNA content. (G) Cyclin D1 expression in cells in J culture without thymidine treatment. R1 in panel G represents GATA2+ cyclin D1− region. Numbers in A and E-G indicate the percentage of cells in each quadrant.

GATA2 expression in normal progenitor-cell fractions. (A) Cord blood CD34+ cells (1 × 105 cells) were cultured for 1 week in medium containing SCF (S), with/without Flt-3L (FL) or Jagged 1/Fc-Protein G-agarose beads (J). Density plot analysis of GATA2 expression and DNA content in the cord blood CD34+ cell fraction. Vertical lines represent borders of 2N population. Bar graphs represent number of total cells (left) and of GATA2+ cells (right) after 1 week's culture. Mean ± standard error of the mean; n = 4. *P < .05 compared with the culture containing SCF. (B) Immunoblot analysis of the cultured cord blood CD34+ fraction. (−) indicates no thymidine addition; T, treated with thymidine for 24 hours; TR, after release from thymidine treatment; and N, treated with nocodazole for 24 hours. (C-G) Lin fractions of mouse bone marrow cells were cultured with SCF and Jagged 1 (J) or SCF and erythropoietin (E) for 4 days, treated with excess thymidine for additional 24 hours, then released into fresh medium. (C) DNA content histogram. (D) Immunoblot analysis. (E-F) Density plot analysis of GATA1, GATA2, and DNA content. (G) Cyclin D1 expression in cells in J culture without thymidine treatment. R1 in panel G represents GATA2+ cyclin D1 region. Numbers in A and E-G indicate the percentage of cells in each quadrant.

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