Effect of Gfi-1B on the expression of genes involved in erythropoiesis.

CD34⁺ cells were transduced with either Bcl-xLor Gfi-1B and were cultured in the presence of SCF and GM-CSF. In addition, CD34⁺ cells transduced with either empty vector (control) or Gfi-1B were cultured in the presence of SCF, IL-3, GM-CSF, and EPO. With exogenousBcl-xL, a small but significant number of erythroblasts developed even in the absence of EPO (data not shown). On day 10 of culture, glycophorin A⁺ erythroblasts were purified by cell sorting, then subjected to RT-PCR analysis. RT-PCR was performed on normalized cDNA templates. PCR products were electrophoresed on agarose gels and visualized by ethidium bromide staining.