Narciclasine

An anti-BAI1 rabbit polyclonal was generated (Covance, Princeton, NJ) against amino acids 1180–1584 of human BAI1 and affinity purified using the antigen purified as a thrombin-cleaved GST-fusion coupled to UltraLink BioSupport (Thermo Scientific, Waltham, MA). We described this antibody previously (Duman et al., 2013 (link)); BAI1 is only detected by this antibody in Western blots of Cos-7 cells when they are exogenously expressing BAI1. The immunoreactivity of this antibody for neurons is strongly decreased by BAI1 Kd. This antibody was used at 0.001 mg/ml for IHC, ICC, and Western blotting and was reused at least 5X. For immunoprecipitation, 1 µg per 1.3 × 10⁶ neurons in culture was used. Rabbit anti-BAI11 N-terminal polyclonal (H-270, sc-66815), rabbit anti-Bcr polyclonal (N-20, sc-885), mouse anti-Abr monoclonal IgG₁ (24, sc-135821), and mouse monoclonal anti-c-myc IgG₁ (9E10, sc-40) were obtained from Santa Cruz Biotechnology (Dallas, TX) and used at 1:1000 dilution for Westerns and ICC. DMSO (D2650) was obtained from Sigma (St. Louis, MO). Tetradotoxin, narciclasine, D-AP5, and Y-27632 dihydrochloride were obtained from Tocris (Minneapolis, MN). Potassium chloride (P217) was obtained from Fisher Scientific (Pittsburgh, PA).

Narciclasine was purchased from Tocris Bioscience (Minneapolis, MN, USA). The cytometric bead array (CBA) Rat Inflammation kit was purchased from BD Biosciences, USA. The rat ELISA kits for TNF-α, IL-6, IL-1β and IFN-γ were purchased from Bioassay Technology Laboratory, China. Rat S100A8 and S100A9 ELISA kits were purchased from CusaBio (China). Antibodies against S100A9, NF-κβp65, IL-6 were purchased from Novus Biologicals (Littleton, CO, USA). Antibodies against S100A8 was purchased from Abcam (Cambridge, MA, USA). Antibodies against TNF-α, β-actin, MyD88, IRAK-1 and TRAF-6 were purchased from ABclonal (Woburn, MA, USA). Antibodies against phospho-NFκβp65, Iκβα were purchased from Cell Signaling technology (Danvers, MA, USA).