.The demethylation of the
-P2 region increases luciferase activity. Results of luciferase assays in transiently transfected HEK293 cells. Forty-eight hours after transfection, basal activity of unmethylated and methylated, Firefly luciferase reporter plasmids containing IGF1 promoter 2 CG fragment, and empty Firefly luciferase reporter plasmid, pCpGL-Basic, was measured, and normalized to the activity of co-transfected Renilla luciferase plasmid. Results were analyzed by paired t-test, **P < 10−4, and bars represent three independent experiments (mean ± SEM).