Supplementary Figures S1-S8 and Supplementary Tables S1-S7_Goldfarb et al_Endocrinology (2022)_Interplay between sex-biased GH-regulated mouse liver transcriptome

Supplementary Figures S1-S8 and Supplementary Tables S1-S7, from a study entitled: <br> Interplay between sex-biased GH-regulated mouse liver transcriptome and hepatic zonation revealed by single nucleus RNAseq <br> [Goldfarb et al, Endocrinology (2022)] <br> Abstract: The zonation of liver metabolic processes is well-characterized; however, little is known about the cell type-specificity and zonation of sexually dimorphic gene expression or its GH-dependent transcriptional regulators. We address these issues using single nucleus RNA-sequencing of 32,000 nuclei representing nine major liver cell types. Nuclei were extracted from livers from adult male and female mice, from males infused with GH continuously, mimicking the female plasma GH pattern, and from mice exposed to TCPOBOP, a xenobiotic agonist ligand of the nuclear receptor CAR that perturbs sex-biased gene expression. Analysis of these rich transcriptomic datasets revealed: 1) expression of sex-biased genes and their GH-dependent transcriptional regulators is primarily restricted to hepatocytes and is not a feature of liver non-parenchymal cells; 2) many sex-biased transcripts show sex-dependent zonation within the liver lobule; 3) gene expression is substantially feminized in both periportal and pericentral hepatocytes when male mice are infused with GH continuously; 4) sequencing nuclei increases the sensitivity for detecting thousands of nuclear-enriched lncRNAs and enables determination of their liver cell type-specificity, sex-bias and hepatocyte zonation profiles; 5) the periportal to pericentral hepatocyte cell ratio is significantly higher in male than female liver; and 6) TCPOBOP exposure disrupts both sex-specific gene expression and hepatocyte zonation within the liver lobule. These findings highlight the complex interconnections between hepatic sexual dimorphism and zonation at the single cell level and reveal how endogenous hormones and foreign chemical exposure can alter these interactions across the liver lobule with large effects on both protein-coding genes and lncRNAs.