In each experiment, at least four microscopic fields were counted. cells exposed to OGD, on the contrary, UPF-1069 (1C10 molL?1) significantly reduced post-ischaemic damage. Conclusion and implications: Selective PARP-2 inhibitors increased post-OGD cell death in a model characterized by loss of neurons through a caspase-dependent, apoptosis-like process (hippocampal slice cultures), but they reduced post-OGD damage and increased cell survival in a model characterized by a necrosis-like process (cortical neurons). UPF-1069 may be a valuable tool to explore the function of PARP-2 in biological systems and to examine the different functions of PARP isoenzymes in the mechanisms of cell death and survival. model of the hippocampal damage common of transient global ischaemia (Moroni for 5 min at 4C. The crude nuclear pellet was washed and resuspended in 1 mL of PARP assay buffer (5 Arbidol mmolL?1 MgCl2, 2 mmolL?1 DTT, 50 mmolL?1 Tris, pH 8) containing 100 molL?1 N-methyl-N-nitro-N-nitrosoguanidine (MNNG) to fully activate PARP activity. Samples made up of 100 L of the resuspended nuclear pellet were incubated for 60 min at 37C in the presence of 35.5 nmolL?13H-NAD. The reaction was stopped with 1 mL of 10% trichloroacetic acid (w/v), and the mixture was centrifuged at 12 000for 10 min at 4C. The reaction was terminated by the addition of 1 mL of 10% trichloroacetic acid (w/v), and radioactivity of the suspension was measured by liquid scintillation spectrometry. Evaluation of tankyrase-1 function HeLa cells cultured in Dulbecco’s altered Eagle’s medium (DMEM) made up of 10% heat-inactivated fetal calf serum were synchronized in mitosis by using 700 nmolL?1 S-trityl-L-cysteine, fixed in paraformaldehyde 4% and processed for immunocytochemical evaluation using turbulent antibodies as described by Chang (2005). In order to reduce the synthesis and function of tankyrase-1, cells were transfected with small interference RNA (siRNA) (control siRNA: 5-AATTCTCCGAACGTGTCACGT, tankyrase-1 siRNA: 5-AACAAUUCACCGUCGUCCUCU, Dharmacon, Lafayette, CO, USA) by using oligofectamine (Invitrogen, San Giuliano Milanese, Italy) as described by the manufacturer, and assayed 2 days post transfection. Imaging was performed by using a Nikon fluorescence microscope equipped with piezoelectric motorization and a CCD camera. Stacks of images were acquired through the depth of the section by sing Metamorph/Metafluor software (Molecular Devices, Downingtown, PA, USA) and deconvoluted by using Image Autodeblur software (MediaCybernetics, Bethesda, MD, USA). For each field, the number of mitosis and the ratio between Arbidol abnormal and normal mitosis were evaluated. In each experiment, at least four microscopic fields were counted. The final values represent the mean of at least three impartial experiments. OGD in rat organotypic hippocampal slices All animal care and the experimental procedures were formally approved by the ethical committee for animal care at the Department of Pharmacology of the University of Florence and were performed in compliance with the recommendations of Arbidol the European Union (86/609/EEC). Organotypic hippocampal slice cultures were prepared as previously described (Pellegrini-Giampietro < 0.01 versus respective control. CRL, control; MNNG, N-methyl-N-nitro-N-nitrosoguanidine; PARP, poly(ADP-ribose) polymerase; TIQ-A, thieno[2,3-< 0.01 versus control; Scale bar: 5 m. CRL, control; PARP, poly(ADP-ribose) polymerase; siRNA, small interference RNA; Arbidol TIQ-A, thieno[2,3-(Kirino, 1982; Pulsinelli < 0.05 versus 20 min OGD; Scale bar: 2 mm. Tmem1 CRL, control; OGD, oxygen-glucose deprivation; PARP, poly(ADP-ribose) polymerase; Arbidol PI, propidium iodide; TIQ-A, thieno[2,3-< 0.05 versus 60 min OGD. Scale bar: 50 m. CRL, control; LDH, lactate dehydrogenase; OGD, oxygen-glucose deprivation; PARP, poly(ADP-ribose) polymerase; TIQ-A, thieno[2,3-(global forebrain ischaemia of 20C30 min) suggest that PARP inhibition reduces the hippocampal damage mostly because of a decreased inflammatory cell infiltration (Hamby showing that these animals have a reduced brain infarct after middle cerebral.