Fig. 13.
Fig. 13. (A) Percentage of cells in the G0 and G1 phases after stimulation with the increasing concentrations of GM-CSF (0.1 to 100 ng/mL) in the presence of 10 U/mL of EPO. Growth factor-deprived UT-7/GM cells were cultured under various conditions and obtained for the analysis of the cell cycle. (B) Effect of the increasing concentration of GM-CSF on the proliferation of UT-7/GM cells in the presence of a constant concentration of EPO. Cell proliferation of UT-7/GM was analyzed by MTT assay (see Materials and Methods). Cells were plated at a density of 104/well in IMDM supplemented with 5% FCS and cultured with increasing concentrations of GM-CSF (0.01 to 10 ng/mL) in the presence of 10 U/mL of EPO. MTT reduction was measured after 3 days of culture. The values represent the mean ± SD from triplicate cultures. (C) Suppression by GM-CSF of EPO-induced erythroid differentiation in a dose-dependent manner. UT-7/GM cells were cultured with increasing concentrations of GM-CSF (0.01 to 10 ng/mL) in the presence of 10 U/mL of EPO. Two weeks later, the cells were obtained and stained with dianisidine as described in Materials and Methods. (D) Percentage of dianisidine-positive cells after stimulation with increasing concentrations of EPO. Cells were cultured with increasing concentrations of EPO (0.1 to 100 U/mL). Two weeks later, the cells were obtained and stained with dianidisine. (E) Percentage of cells in the G0 and G1 phases after stimulation with increasing concentrations of EPO. Growth-factor–deprived cells were cultured with increasing concentrations of EPO (0.1 to 100 U/mL) for the indicated periods and then harvested for the analysis of the cell cycle.

(A) Percentage of cells in the G0 and G1 phases after stimulation with the increasing concentrations of GM-CSF (0.1 to 100 ng/mL) in the presence of 10 U/mL of EPO. Growth factor-deprived UT-7/GM cells were cultured under various conditions and obtained for the analysis of the cell cycle. (B) Effect of the increasing concentration of GM-CSF on the proliferation of UT-7/GM cells in the presence of a constant concentration of EPO. Cell proliferation of UT-7/GM was analyzed by MTT assay (see Materials and Methods). Cells were plated at a density of 104/well in IMDM supplemented with 5% FCS and cultured with increasing concentrations of GM-CSF (0.01 to 10 ng/mL) in the presence of 10 U/mL of EPO. MTT reduction was measured after 3 days of culture. The values represent the mean ± SD from triplicate cultures. (C) Suppression by GM-CSF of EPO-induced erythroid differentiation in a dose-dependent manner. UT-7/GM cells were cultured with increasing concentrations of GM-CSF (0.01 to 10 ng/mL) in the presence of 10 U/mL of EPO. Two weeks later, the cells were obtained and stained with dianisidine as described in Materials and Methods. (D) Percentage of dianisidine-positive cells after stimulation with increasing concentrations of EPO. Cells were cultured with increasing concentrations of EPO (0.1 to 100 U/mL). Two weeks later, the cells were obtained and stained with dianidisine. (E) Percentage of cells in the G0 and G1 phases after stimulation with increasing concentrations of EPO. Growth-factor–deprived cells were cultured with increasing concentrations of EPO (0.1 to 100 U/mL) for the indicated periods and then harvested for the analysis of the cell cycle.

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