Figure 3

The Oct-1 decoy oligonucleotide-induced expression of the endogenous γ -globin genes in K562 cells. The K562 cells were treated with either the decoy or the scrambled oligonucleotides at indicated concentrations. As a positive control, some K562 cells were treated with 75 μM hemin. Total RNA was isolated from both untreated and treated cells at various time points following the treatment. The level of γ-globin mRNA was determined by a reverse transcription-based quantitative PCR (real time PCR). In order to determine the level of HbF in the treated K562 cells, the cells were treated with the decoy or the scrambled oligonucleotide for four days and then harvested and analyzed by western blots using an antibody that recognized the HbF. (A) The Oct-1 decoy oligonucleotide treatment-induced γ-globin transcription as determined by real time PCR assay. The level of the γ-globin mRNA in the untreated K562 cells was counted as 100% and the levels of the γ-globin mRNA in the treated K562 cells were calculated as relative levels to that of the untreated K562 cells. The results are from at least three independent experiments. (B) The Oct-1 decoy oligonucleotide treatment-induced accumulation of HbF in the K562 cells. The results are from three individual experiments. * Statistical significance between the untreated and the treated K562 cells at the same time point with p < 0.01.