FIG. 3.

Effect of prenatal cocaine exposure on CpG methylation in the Prkce promoter in the left ventricle. Time-dated pregnant Sprague-Dawley rats were administered saline or cocaine (30 mg/kg/day) from Days 15 to 20 of gestational age, and left ventricles were obtained from 3-mo-old male (m) and female (f) offspring. DNA was extracted and treated with sodium bisulfite. The modified DNA was used as a template for real-time fluorogenic methylation-specific PCR. Primers for the Prkce promoter regions of interest are listed in Table 1. Gapdh was used as an internal reference gene. Real-time methylation-specific PCR was performed using the iQ SYBR Green Supermix with iCycler real-time PCR system. Data are mean ± SEM, n = 6. Data were analyzed by two-way ANOVA for each binding site, with cocaine exposure as one factor and gender as the other. The underlining indicates the binding sequences. *P < 0.05, cocaine vs. control.