Iem 1460

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.