Insulin

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)

Cell culture media were obtained from PAA (Coelbe, FRG). Recombinant human (rh) interleukin (IL) types 1β IΛ-1β and rh-TNFα were purchased from Endogene (MA, USA). A commercial enhanced chemiluminescence detection kit with nitrocellulose membranes (Hybond-C) was obtained from Amersham (Braunschweig, FRG). Diclofenac and bovine serum albumin were from Sigma (Deisenhofen, FRG). Primers were synthesized by MWG-Biotech (Ebersberg, FRG); AmpliTaq polymerase was obtained from Perkin-Elmer (Weiterstadt, FRG) and SuperScript reverse transcriptase from GIBCO (Eggenstein, FRG). Insulin, transferrin supplements, and recombinant human interferon (IFN) type-γ rh-IFN-γ were from Boehringer (Mannheim, FRG). Primary antibodies to characterize cellular cultures were obtained from Dako (CA, USA); secondary antibodies were from Dako and Dianova (Hamburg, FRG).