We used patch methylation to examine whether selective methylation at specific sites reduces NGFI-A binding to and transactivation through the exon 1F NR3C1 promoter. We found an effect of methylation status on transcription factor–induced gene expression from the NR3C1 promoter (Fig. 3c). For the 125-bp construct, there was a significant effect of methylation status (F = 57.6, P < 0.0001) and NGFI-A treatment (F = 6.3, P < 0.05). As predicted, there was also a significant interaction between methylation status and the NGFI-A expression (F = 48.7, P < 0.0001). Post hoc analysis of the 125-bp NR3C1 promoter construct revealed that the effect of NGFI-A on gene transcription was significantly (P < 0.001) greater in the presence of the unmethylated rather than the patch-methylated NR3C1 promoter construct. The same pattern of results was observed for the 255-bp NR3C1 promoter construct. Thus, for the 255-bp NR3C1 promoter construct, there was a significant effect on transcriptional activity of methylation status (F = 555.4, P < 0.0001) and NGFI-A expression (F = 387.3, P < 0.0001). There was also a significant interaction between
