Figure 6.

Exposure to the PFAS mixture, hepatic cholesterol and bile acid levels, and synthesis pathways. (A) Hepatic protein was isolated from $n=5$ mice from each treatment group and hepatic total cholesterol levels were measured. (B) Total hepatic RNA ($n=10$ mice from each treatment group) was isolated, and gene expression levels of Hmgcr were determined by RT-qPCR. (C) Diagram of hepatic cholesterol and bile acid synthesis pathways. Steps preceded by “&” represent a significant reduction, whereas those measured and not significantly affected are preceded by “@.” (D) Hepatic total bile acid levels. Gene expression levels of (E) Cyp7a1, and (F) Cyp27a1 were determined by RT-PCR. GAPDH was used as a housekeeping gene. Two-way ANOVA was used to analyze both main effects (i.e., sex, PFAS), as well as the interaction between sex and PFAS (interaction $p<0.05$). The Holm–Sidak post hoc test was used for multiple comparisons. With past consultation from biostatisticians, a significant interaction term supersedes the main effects and can make their meaning unclear. We therefore have not included the main effects $p$-values for any figure with a significant interaction. Box plots represent the median values with upper and lower quartiles; whiskers extend to the 1st and 99th percentiles. Data are reported in Excel Tables S12 and S13. Note: ANOVA, analysis of variance; GAPDH, glyceraldehyde 3-phosphate dehydrogenase; HMG-CoA, 3-hydroxy-3-methylglutaryl coenzyme A; PFAS, per- and polyfluoroalkyl substances; RT-qPCR, real-time quantitative polymerase chain reaction.