Sex-biased genetic programs in liver metabolism and liver fibrosis are controlled by EZH1 and EZH2 (original) (raw)

liver. Hepatic Ezh1/Ezh2 deficiency is also shown to down regulate many male-biased genes, presumed as a secondary response to the disruption of female-biased gene expression. Finally, we show that many genes associated with liver fibrosis and liver carcinogenesis are differentially responsive to the loss of Ezh1/Ezh2 in male compared to female liver, which may contribute to the observed sex-differences in the incidence and progression of liver cancer. Methods Animal tissues. Livers from 7-week-old male and female Ezh1-knockout mice with a hepatocytespecific knockout of Ezh2 (E1/E2-KO mice, also designated Double-knockout (DKO) in the figures and tables) and their age and sex matched floxed littermate controls were generated as described [33]. Briefly, Ezh2 fl/fl mice [40] were bred with Alb-Cre transgenic mice [41]; their offspring were then bred with Ezh1-knockout mice (Thomas Jenuwein, Research Institute of Molecular Pathology, Vienna, Austria) [42] to generate E1/E2-KO mice. Livers from 2-8 week old male and female CD1 mice (ICR strain) were those described previously [43]. Hypophysectomy and continuous GH infusion of male mice for 14 d using an Alzet osmotic minipump were performed as described [19, 44]. Livers used in this study were obtained from mice housed and handled according to NIH guidelines, and all animal experiments were approved by the Animal Care and Use Committee of National Institute of Diabetes and Digestive and Kidney Diseases. qPCR analysis. Liver total RNA (1 μg) was reverse transcribed using the Applied Biosystems High-Capacity cDNA Reverse Transcription Kit (Fisher, Cat#43-688-14). qPCR was performed using Power SYBR green PCR master mix and processed on an ABS 7900HT sequence detection system (Applied Biosystems) or the CFX384 Touch Real-Time PCR detection system (Bio-Rad). For RT-qPCR, raw Ct values were analyzed using the comparative Ct method with normalization to the 18S RNA content of each sample. Primers used for qPCR are shown in Table S1A.