FIG. 3.

Mutational analyses of the rat ANF promoter-enhancer. (A) Schematic diagram of the ANF enhancer-promoter showing the positions of the two A/T-rich sequences (A/T) that JMJ binds to, Nkx2.5-binding sites (N), and GATA4-binding sites (G). (B) Mutational analyses of the ANF reporter genes. The mutant ANF reporter genes (2 μg) were cotransfected with Nkx2.5 (1 μg) and GATA4 (1 μg) and/or JMJ (1 μg) expression vector into 10T1/2 cells as described for Fig. 1B. Filled bars represent the means and T bars indicate the standard errors of the means for three separate transfection assays with duplicate plates. *, P < 0.05; **, P < 0.01 compared to respective control activation.