Fig. 4
Comparison of the hepatic transcriptome between the adult offspring of ethanol-exposed and control males. a Volcano plot displaying differences in gene expression between adult male offspring sired by ethanol-exposed and control fathers (n = 3, cutoff q = 0.05). b Heatmap depicting the differentially expressed genes between the pooled control offspring and each of the three offspring of the alcohol-exposed males. c Top three categories identified using Ingenuity Pathway Analysis. d RT-qPCR validation of candidate genes related to the identified FXR/RXR, FXR/RXR and cholesterol biosynthesis pathways in the adult male liver (n = 8). Levels of hepatic e total cholesterol, f high-density lipoprotein and g low-density lipoprotein cholesterol esters within the liver of male and female offspring sired by ethanol-exposed and control fathers (n = 8). h Proportional ratios of HDL cholesterol to total cholesterol and LDL cholesterol to total cholesterol in liver samples derived from the male and female offspring of ethanol-exposed and control males (n = 8). Quantification of i total cholesterol, j high-density lipoprotein and k low-density lipoprotein cholesterol esters within the plasma of male and female offspring sired by ethanol-exposed and control fathers (n = 8). l Proportional ratios of HDL cholesterol to total cholesterol and LDL cholesterol to total cholesterol in plasma derived from the male and female offspring of ethanol-exposed and control males (n = 8). For RT-qPCR analyses, measured Ct values were normalized to the geometric mean of transcripts encoding Ywhaz, Hprt and Mrpl1, and graphed relative to the control treatment. Graphs represent independent replicates, with two independent RT reactions and three RT-qPCR measurements for each RT. Data were analyzed using either an unpaired t test, arcsine transformed and an unpaired t test with Welch’s correction applied or used in a two-way ANOVA followed by Sidak post hoc analysis. Error bars represent SEM *p < 0.05 and **p < 0.01 (comparisons between alcohol and control preconception treatments)