Leupeptin hemisulfate

Dulbecco’s Modified Eagle’s Medium (DMEM) (#11995065), Neurobasal Medium (#21103049), trypsin-EDTA 0.25% (#25200056), penicillin-streptomycin (#15140122), B27 Supplement (#17504944), and Glutamax (#35050061) were from Life Technologies (Carlsbad, CA, USA), Fetal calf serum (FCS) was from Internegocios SA (Mercedes, Argentina). The protease inhibitors, purchased from Santa Cruz Biotechnology (Dallas, TX, USA), were leupeptin hemisulfate (#295358), pepstatin A (#45036), aprotinin (#3595), and phenylmethylsulfonyl fluoride (#329-98-6). Monosialoganglioside GM2 (#1502) was obtained from Matreya (State College, PA, USA). The primary antibodies used were anti-P-PERK (#3179), anti-CHOP (#L63F7) (Cell Signaling Technology, Danvers, MA, USA), anti-CHOP (#MA1-250) (Thermo Fisher, Waltham, MA, USA), anti-MAP-2 (#PCK-554P) (Covance Inc. Princeton, NJ, USA), and anti-MAP-2 (#M2320) (Sigma, St. Louis, MO, USA).

Histopaque, p-nitrophenyl valerate (pNPVa), and LPS (E. coli 055:B5) were purchased from Sigma (St. Louis, MO, USA). AIM V^® Serum Free Medium was purchased from Thermo-Fisher (Waltham, MA, USA). RIPA lysis buffer and protease inhibitors (phenylmethylsulfonyl fluoride, PMSF; 4-(2-aminoethyl)benzenesulfonyl fluoride, AEBSF; bestatin; pepstatin A; leupeptin hemisulfate; and aprotinin) were from Santa Cruz Biotechnology (Dallas, TX, USA). Antibodies used for Western blots (anti-CES1, anti-MAGL, β-actin, goat anti-rabbit, and goat anti-mouse) were purchased from Abcam (Cambridge, MA, USA). Authentic 2-AG, anandamide, AA, and its deuterated analog AA-d8 were purchased from Cayman Chemical Company (Ann Arbor, MI, USA). Small molecules JZL184 and WWL113 were purchased from Sigma. CpG was purchased from InvivoGen (San Diego, CA, USA). Primary CES1 and MAGL antibodies for flow cytometry were purchased from Abcam. Fluorescence secondary antibodies for flow cytometry and antibodies against IL-6 used to neutralize IL-6 or perform ELISA were purchased from Biolegend (San Diego, CA, USA). For some of the experiments, monocyte-depleted (n = 1) and whole PBMCs (n = 5) were purchased from Astarte Biologics (Bothell, WA, USA).