Xylacin

For the described experiments the following materials and chemicals were used: Minimal essential medium (MEM), glutamine, Hanks' Balanced Salt Solution (HBSS), normal horse serum (NHS; Gibco BRL Life Technologies, Eggenstein, Germany), glucose (Braun, Melsungen, Germany), cell-culture inserts with a pore size of 0.4 µm (Millipore, Schwalbach/Ts., Germany), vibratome (Vibratom VT 1200 S; Leica Microsystems AG, Wetzlar, Germany), six-well culture dishes (Falcon, BD Biosciences Discovery Labware, Bedford, MA, USA), streptomycin, penicillin, ascorbic acid, N-methyl-D-aspartate (NMDA; Sigma-Aldrich, Deisenhofen, Germany), insulin (Boehringer, Mannheim, Germany), ketamine hydrochloride (Merial, Hallbergmoos, Germany), xylacin (Bayer, Leverkusen, Germany, atropine (Ratiopharm, Ulm, Germany), bicinchoninic acid (BCA) test, Tris (Roth, Karlsruhe, Germany), Glycerol (Sigma-Aldrich), β-mercaptoethanol (Sigma-Aldrich, Steinheim, Germany), bromophenol blue (AppliChem, Darmstadt, Germany), chemiluminescence, superfrost microscope slides from (Thermo Fisher Scientific, Rockford, IL, USA), cryostat 3050 S (Leica), coated slides (Menzel, Braunschweig, Germany), X-ray films (Kodak, Stuttgart, Germany), Dako fluorescent mounting medium (Dako, Hamburg, Germany), Entellan (Merck, Darmstadt, Germany)

At an age of thirteen weeks, brains from 8 male and 9 female mice prenatally exposed to Poly I:C, and 9 male and 9 female offspring from PBS-treated dams were used for epigenetic analysis. For this purpose, mice were anesthetized with a single intraperitoneal injection of Ketamin (Pfizer, Germany) and Xylacin (Bayer, Germany). 0.7 ml 10 % Ketamin and 0.3 ml 2 % Xylacin were mixed and administered at a dose of 10 ml/kg. Afterwards, brains were collected and stored at −20 °C in RNA-Later (Invitrogen, Germany). Half of the brain was used to extract nuclear proteins using a Nuclear Extract Kit (Active Motif, Belgium) and the activities of various HDAC and DNMT enzymes were analyzed with the HDAC Fluorescent Assay Kit and DNMT Activity/Inhibition Assay (Active Motif, Belgium). The fluorescence/optical density was detected with a PolarStar OPTIMA microplate fluorimeter (BMG Labtech).