title: Assessing the Histone Deacetylase Activity of SIRT6 in Primary Murine Hepatocytes via Proximity Ligation Assay.
abstract: Generation of primary cell culture of hepatocytes by mouse liver perfusion (MLP) combines the advantages of in vivo and in vitro models. It provides hepatocytes that grow under physiological conditions in mice, with the genotype of the whole organism or a specific gene knockout. In contrast to immortalized cell cultures, primary murine hepatocytes (pmHep) are non-cancerous cells with a limited lifespan but still amenable to classical in vitro methods such as treatment with drugs, small molecule inhibitors, and agonistic/antagonistic antibodies of surface receptors as well as transfection. One technique, which has gained popularity recently, is the analysis of protein-protein interactions by the proximity ligation assay (PLA). Here, we describe a liver perfusion protocol and the detection of the histone deacetylase function of Sirtuin 6 (SIRT6) using PLA in pmHep.
authors.affiliation: First Department of Internal Medicine, Molecular Hepatology, University Medical Center, Johannes Gutenberg University Mainz, Obere Zahlbacherstr Straße 63, 55131, Mainz, Germany.
authors.forename: Katharina
authors.initials: K
authors.lastname: Wolf
authors.affiliation: First Department of Internal Medicine, Molecular Hepatology, University Medical Center, Johannes Gutenberg University Mainz, Obere Zahlbacherstr Straße 63, 55131, Mainz, Germany. sstrand@uni-mainz.de.
authors.forename: Susanne
authors.initials: S
authors.lastname: Strand
mesh_terms: D000818:Animals; D001681:Biological Assay; D018929:Cell Culture Techniques; D022781:Hepatocytes / Q000166:cytology / Q000201:enzymology*; D008099:Liver / Q000166:cytology / Q000201:enzymology; D051379:Mice; D018345:Mice, Knockout; D010477:Perfusion / Q000379:methods; D061251:Primary Cell Culture; D011485:Protein Binding; D025941:Protein Interaction Mapping / Q000379:methods*; D037761:Sirtuins / Q000172:deficiency / Q000235:genetics*
chemical_list: C501860:Sirt6 protein, mouse; D037761:Sirtuins
keywords: Histone deacetylase; Mouse liver perfusion; Primary murine hepatocytes; Protein–protein interactions; Proximity ligation assay; Sirtuin 6