L glutamic acid monosodium salt hydrate

l-Glutamic acid monosodium salt hydrate (Sigma) was dissolved in water and added to the media to reach a final concentration of 500 μM, which is similar to reported in vivo concentrations of glutamate after acute SCI (Xu et al., 2004 (link)). The following inhibitors were used: NBQX (Tocris; 10 μM), a competitive antagonist of AMPAR/KARs, and IEM 1460 (Hello Bio; 30 μM), an open channel CP-AMPAR blocker. The following agonists or positive modulators were used: AMPA (Tocris; 50 μM) and CX546 (Alomone Labs, 50 μM). The inhibition of Notch was achieved with the pharmacological gamma-secretase inhibitor, DAPT (Sigma; 1 μM). To target Ca²⁺-dependent pathways, the following inhibitors were used: W-13 hydrochloride (W-13), Autocamtide-2-related inhibitory peptide (AIP), and 666-15 (Xie et al., 2015 (link)). Stock concentrations of each agent were dissolved in the appropriate solvents (water or DMSO) and applied to media to achieve the final working concentration. The final concentration of inhibitors used for experiments demonstrated no toxicity when administered to epNSPCs alone (Figure S1A). For control conditions, epNSPCs were treated with media plus vehicle alone.

d-glucose, Dimethylsulfoxide (DMSO), hydrochloric acid, Sodium hydroxide, bovine serum albumin, and sodium chloride were purchased from Carl-Roth (Kalshur, Germany). Epinephrine, l-Glutamic acid Monosodium salt hydrate, hexadecyl-trimethylammonium bromide, sodium azide, O-dianisidin Thiobarbituric acid, Trichloro-acetic acid, and paraformaldehyde were purchased from Sigma-Aldrich Chemical Co (Taufkirchen, Germany). Interleukin-1 (IL-1β) and tumor necrosis factor-alpha (TNF-α) kits were purchased for R&D Systems while sodium thiopental, atorvastatin tablet, and Insulin (Actrapid, Novo Nordisk) were purchased from a local pharmacy (Dschang, Cameroon). All chemical reagents used in this study were of analytical grade.

All cell culture media and supplements were procured from Gibco, unless stated otherwise. L-Glutamic acid monosodium salt hydrate was obtained from Sigma-Aldrich, while cordycepin was purchased from Aladdin. KeyGen Biotech supplied the Acridine Orange Detection Kit and Reactive Oxygen Species Assay Kit, while Solarbio provided the Acetylcholinesterase Assay Kit. The GSH kit was procured from Nanjing Jiancheng Bioengineering Institute.