SBH significantly alleviates FAC-induced loss of neuronal cell viability and cytotoxicity. Cell viability (A, C, E) and cytotoxicity (B, D, F) were evaluated in HT22 hippocampal neuronal cells exposed to either SBH alone (0–100 µM; A, B), FAC (75 µM; C, D) or TBHP (200 µM; E, F) for 24 h, in the presence of SBH (10 µM) or DMSO as a vehicle control. Data is presented as mean ± SEM. *p < 0.05, ****p < 0.0001, one- or two-way ANOVA followed by Bonferroni post hoc analysis. n = 12 replicates from 4 independent experiments

SBH attenuates markers of ferroptosis. HT22 hippocampal neurons were exposed to FAC (75 µM) for 24 h, in the presence and absence of SBH (10 µM) or ChB (50 µM). Cell viability was assessed by CCK-8 assay (A). Iron accumulation (B) and lipid peroxidation (C) were determined in HT22 cell protein lysates. The mRNA expression of Gpx4 (D), Hmox1 (E), and Acsl4 (F) was determined as key indicators of ferroptosis. Data is presented as mean ± SEM. ****p < 0.0001, two-way ANOVA and Bonferroni analysis. n = 9 replicates from 3 independent experiments

FAC induces a loss of morphological and behavioural integrity in zebrafish larvae in vivo. Zebrafish larvae (4 dpf) to FAC (0–600 µM) for 24 h. Individual larvae were assessed for survival (A), incidence of non-lethal malformations (B) and touch startle response (C). Data presented as mean ± SEM (A) or as a proportion of total larvae (B, C). Statistical comparisons were made by one-way ANOVA or Chi-squared analysis. **p < 0.01, ***p < 0.001, ****p < 0.0001. n = 45 larvae from 3 independent experiments

SBH significantly protects against FAC-induced toxicity in zebrafish larvae in vivo. Zebrafish larvae (4 dpf) were injected i.v. with SBH (10 µM) or vehicle, in the presence and absence of bath-applied FAC (600 µM; 24 h). Larvae were assessed for survival (A), incidence of non-lethal malformations (B) and touch startle response (C). Data presented as mean ± SEM (A) or as a proportion of total larvae (B, C). Statistical comparisons were made by two-way ANOVA or Chi-squared analysis. **p < 0.01, ***p < 0.001, ****p < 0.0001. n = 45 larvae from 3 independent experiments