FIGURE 3.

PEMT risk allele is not estrogen-responsive. A, RNA isolated from primary hepatocytes heterozygous for the risk allele was subjected to allele-specific quantitative PCR. Results were normalized to genomic DNA (black bars). The estimated relative expression of the risk allele cDNA versus the protective allele cDNA was calculated by linear regression analysis. In the absence of 100 nmol/liter moxestrol treatment (−, open bars), the risk allele was underexpressed by ∼4% relative to the protective allele. Upon moxesterol treatment (24 h) (+, gray bars), the risk allele was underexpressed by ∼8%. Based on the transcript-specific expression data, the estimated relative abundance of unregulated transcript A in treated cells is depicted by the dashed line. Results are presented for individual subjects and as an average for all subjects (n = 9). *, p < 0.01; **, p < 0.001. Error bars, S.E. B, RNA was isolated from moxestrol-treated (open diamonds) or untreated hepatocytes (black squares) that were P, R, or P/R. PEMT levels were determined by qPCR. Transcript A was not induced by estrogen in any of the groups. Transcripts B and C were induced by hormone in the P and P/R groups, but in hepatocytes homozygous for the risk allele (R), these transcripts were not induced by hormone treatment. Results are presented as the natural log (LN) of the ratio of PEMT relative to GAPDH gene expression levels in moxestrol-treated versus untreated hepatocytes. (n = 5–6/genotype). *, p < 0.001; **, p < 0.0001. Error bars, S.E.