Figure 1. Systematic pharmacologic characterization of hemin-induced cell death in primary cortical neurons. Cell death is inhibited by chemical inhibitors of ferroptosis or necroptosis.

(A) Primary neurons treated with 100μM hemin (LD₅₀) and chemical inhibitors effective in distinct cell death models (e.g. apoptosis, autophagy, ferroptosis) were examined. Values show mean ± SD at representative concentration in brackets. Grayscale coding indicates the continuum from no protection in the presence of hemin (black) to maximal cell viability (white). (B) Concentration-responses of inhibitors that inhibit hemin-induced death. Values represent mean ± SD, except for U0126 where medians are given. * p<0.05 versus hemin, # p<0.05 versus U0124. (C) Representative live/dead staining are shown. Scale bar = 50μm. (D) Statistical analysis of profile of chemical inhibitors between operationally defined ferroptosis^{20, 27} and hemin-induced toxicity revealed that hemin toxicity in primary neurons can be considered ferroptotic.