Figure 3
Figure 3. The EKLF-GATA1 fusion proteins enhanced δ-globin gene expression in K562 and CD34+ cells. (A) K562 cells were transfected with EKLF, GATA1, fusion EKLF-GATA1 (EG), or vector only and cultured in medium with 6 μg/mL of blasticidin for 2 weeks. RNA was isolated and real-time PCR analysis performed. Data represent real-time PCR analysis of δ- and γ-globin gene expression in EKLF-, GATA1-, or fusion EKLF-GATA1–transfected cells normalized to β-actin gene expression. Fold increase was calculated compared with expression in mock-transfected cells. *P < .05 versus mock-transfected cells. Error bars indicate SD of the mean of 3 independent experiments. (B) CD34+ bone marrow cells were infected by lentivirus encoding EKLF, GATA1, fusion EKLF-GATA1 (EG) constructs, or vector only at day 4 of the expansion stage, On day 6, gene-transduced CD34+ cells were reseeded and grown in differentiation medium with 3 μg/mL of blasticidin for 7 days. RNA was isolated and real-time PCR analysis performed. Data represent real-time PCR analysis of δ-, γ-, and β-globin gene expression in EKLF-, GATA1-, or fusion EKLF-GATA1–transduced cells normalized to β-actin gene expression. Fold increase was calculated compared with vector only–transduced cells. *P < .05 versus vector only–transduced cells. Error bars indicate SD of the mean of 3 independent experiments.

The EKLF-GATA1 fusion proteins enhanced δ-globin gene expression in K562 and CD34+ cells. (A) K562 cells were transfected with EKLF, GATA1, fusion EKLF-GATA1 (EG), or vector only and cultured in medium with 6 μg/mL of blasticidin for 2 weeks. RNA was isolated and real-time PCR analysis performed. Data represent real-time PCR analysis of δ- and γ-globin gene expression in EKLF-, GATA1-, or fusion EKLF-GATA1–transfected cells normalized to β-actin gene expression. Fold increase was calculated compared with expression in mock-transfected cells. *P < .05 versus mock-transfected cells. Error bars indicate SD of the mean of 3 independent experiments. (B) CD34+ bone marrow cells were infected by lentivirus encoding EKLF, GATA1, fusion EKLF-GATA1 (EG) constructs, or vector only at day 4 of the expansion stage, On day 6, gene-transduced CD34+ cells were reseeded and grown in differentiation medium with 3 μg/mL of blasticidin for 7 days. RNA was isolated and real-time PCR analysis performed. Data represent real-time PCR analysis of δ-, γ-, and β-globin gene expression in EKLF-, GATA1-, or fusion EKLF-GATA1–transduced cells normalized to β-actin gene expression. Fold increase was calculated compared with vector only–transduced cells. *P < .05 versus vector only–transduced cells. Error bars indicate SD of the mean of 3 independent experiments.

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