β nicotinamide adenine dinucleotide hydrate n7004

C57BL/6 wild-type mice obtained from Janvier Labs were used for all experiments. Mice were housed in a specific pathogen-free facility and were aged of 8 weeks at the beginning of experiments. All animal experiments were approved by the local institutional ethic committee.
Adenosine 5’-tri-phosphate disodium salt (A2383) and β-nicotinamide adenine dinucleotide hydrate (N7004) were purchased from Sigma Aldrich. Red blood cell (RBC) lysis/fixation Solution, True-Nuclear transcription factor buffer set, fluorochrome-conjugated streptavidin, and antibodies to CD45 (clone 30-F11), CD4 (RM4-5), CD8 (53-6.7), CD25 (PC-61), CD19 (1D3/CD19), B220 (RA3-6B2), FoxP3 (MF-14), CD27 (LG.3A10), CD62L (MEL-14), CD69 (H1.2F3) or P2X7R (1F11), and purified CD16/CD32 (TruStain FcX) were obtained from Biolegend or Sony Biotechnology. Rabbit polyclonal antibody K1G, specific to mouse P2X7, was described in our previous studies (12 (link), 13 (link)). K1G was used to stain P2X7 at the surface of blood myeloid cells as illustrated in Supplemental Figure 3, using a secondary donkey anti-rabbit IgG from Jackson ImmunoResearch. Biotinylated polyclonal antibody specific to mouse IgGa was obtained from Jackson ImmunoResearch and monoclonal antibody to ARTC2.2 (Nika102) from Novus Biologicals.

Normal saline containing 1 mM CaCl₂ was the control solution. Ruthenium Red, DL-2-amino-5-phosphonopentanoic acid, (+)-MK-801 hydrogen maleate, and Memantine hydrochloride (all from Sigma, St. Louis, MO) were diluted, respectively, to 30 μM, 100 μM and 50 μM. Ketamine hydrochloride (Zoetis) was diluted to 50 μM. Glutamate and Glycine (Sigma, St. Louis, MO) were diluted to 1 mM in saline. The enantiomeric forms of GsMTx4 were made and purified according to previously published protocols55 . L-Glutamic Dehydrogenase from Bovine Liver (G2626) and β-Nicotinamide adenine dinucleotide hydrate (N7004) (Sigma, St. Louis, MO) were diluted respectively to 56 U/ml and 1 mM in saline and pre-incubated with the cells before the glutamate assay25 (link).