Fig. 5.

Ferrostatin-1 inhibits glutamate-induced oxytosis and preserves mitochondrial integrity. a: MTT assay revealed protection of BI-6c9 (10 µM) and ferrostatin-1 (2 µM) against glutamate (4 mM, 16 h) toxicity (n=8). b: Cells were stained with BODIPY 581/591 and lipid peroxides were measured by FACS analysis after 16 h of glutamate treatment (3 mM). Ferrostatin-1 significantly reduced the lipid peroxide production (n=4). c: Glutamate treatment (3 mM, 16 h) increased mitochondrial ROS production, which was fully blocked by co-treatment with 2 µM ferrostatin-1 (n=4). d: Quantification of mitochondrial morphology in cells exposed to glutamate showed an increase in cells with mitochondria of category III. This glutamate-induced fragmentation was fully prevented by ferrostatin-1. ##p<0.01 compared to Cat I of glutamate-treated control; ***p<0.001 compared to Cat III of glutamate-treated cells (ANOVA, Scheffé’s test). Mean values were pooled from 4 independent experiments, where mitochondrial morphology was determined from at least 500 cells per condition without knowledge of treatment history. e: Quantification of TMRE fluorescence of n=4 independent experiments showed that MMP was fully restored by ferrostatin-1 (2 µM) after glutamate exposure (3 mM, 16 h). f: After 16 h of treatment with glutamate (3 mM) ATP levels were measured. Ferrostatin-1 prevented ATP depletion compared to glutamate-treated controls (n=8). All results are given as mean + S.D. ###p<0.001 compared to untreated control; ***p<0.001 compared to glutamate-treated control (ANOVA, Scheffé‘s test).