Methylation of α^π-promoted reporter vector (α^πpGL3E) represses transcription.

Primary erythroid cells were transfected with the (A) unmethylated and fully methylated α^πpGL3E vectors or (B) α^πpGL3E vector in which only the α^πpromoter has been methylated or mock methylated. Cells were transfected with 2 μg expression vector. After 48 hours' growth in complete medium, cell lysates were prepared and assayed for firefly luciferase activity as described in “Materials and methods.” The figure shows the relative luciferase activity of the unmethylated and methylated vectors. The values shown are normalized to pRL-TK as a control for transfection efficiency. The error bars represent the SEM for measurements of 3 different samples.