5-Azacytidine increases nascent mRNA production from the γ, β, and α-globin genes. Quantitative RT-PCR was performed using primers that crossed intron-exon boundaries or were within an intron to determine levels of nascent or pre-mRNA as an indicator of gene activity. (A) Effect of 5-azacytidine on nascent γ-globin gene expression. (B) Effect of 5-azacytidine on nascent β-globin gene expression. (C) Effect of 5-azacytidine on nascent α-globin gene expression. (D) Effect of 5-azacytidine on nascent GAPDH gene expression. CD34⁺ cells from a single normal donor were used for this experiment. Each RT-PCR analysis was performed in triplicate.