Monosodium l glutamate

5 × 10³ cells per well were seeded into sterile 96-well plates and incubated
overnight. For the neurotoxicity assay, medium was removed, and increasing
concentrations of the compound diluted with medium from a 0.1 M stock
solution were added to the wells. DMSO (0.05%) in DMEM served as control.
Cells were incubated for 24 h if neurotoxicity was determined by using
a colorimetric MTT assay. For the oxytosis assay, 5 mM glutamate (monosodium l-glutamate, Sigma-Aldrich, Munich, Germany) together with increasing
concentrations of the respective compounds was added to the cells
and incubated for 24 h. As a positive control, a mixture of 25 mM
quercetin (Sigma-Aldrich, Munich, Germany) and 5 mM glutamate was
used. After 24 h incubation, cell viability was determined by using
a colorimetric MTT assay, as described above. Results are presented
as percentage of untreated control cells. Data are expressed as means
± SEM of three independent experiments. Analysis was accomplished
by using GraphPad Prism 9 software by applying one-way ANOVA followed
by Dunnett’s multiple comparison post-test. Levels of significance:
*p < 0.05; **p < 0.01; and
***p < 0.001.

HT22 cells were seeded into sterile 96‐well plates at a density of 5×10³ per well and incubated overnight. The medium was exchanged, and cells were treated with 5 mM glutamate (monosodium‐L‐glutamate, Sigma Aldrich, Munich, Germany) alone or together with 1.56, 3.12, 6.25 or 12.5 μM of the respective compound. After 24 hours cell viability was determined using a colorimetric MTT assay as described above. Results are presented as percentage of untreated control cells. Data is expressed as means ± SEM of three independent biological experiments. Each biological experiment contained three technical replicates. Analysis was accomplished using GraphPad Prism 5 Software applying Oneway ANOVA followed by Dunnett's multiple comparison posttest. Levels of significance: * p<0.05; ** p<0.01; *** p<0.001.

HT22 cells were seeded into sterile black walled 96‐well plates at a density of 5×10³ per well and incubated overnight. The medium was exchanged, and cells were treated with 5 mM glutamate (monosodium‐L‐glutamate, Sigma Aldrich, Munich, Germany) alone or together with 1.56, 3.12, 6.25 or 12.5 μM of the respective compound for 6 hours. The medium was removed and 100 μL phenol red‐free Hank's balanced salt solution (Sigma Aldrich, Munich, Germany) containing 1 μM CM‐H₂DCFDA (Thermo Fisher Scientific, Darmstadt, Germany) was added. After 20 min incubation, fluorescence (λ excitation=495 nm, λ emission=525 nM) was determined using a Tecan Infinite M Plus microplate reader or subjected to fluorescence microscopy using a Zeiss Axiovert Observer fluorescence microscope. Fluorescence was normalized to control cells not exposed to glutamate. Images were processed with the ZEN 3.4 (blue edition) software. Data is expressed as means ± SEM of three independent biological experiments. Each biological experiment contained three technical replicates. Analysis was accomplished using GraphPad Prism 5 Software applying Oneway ANOVA followed by Dunnett's multiple comparison posttest. Levels of significance: */# p<0.05; **/## p<0.01; ***/### p<0.001.