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. 2022 Jul 29;13(7):662. doi: 10.1038/s41419-022-05121-z

Fig. 2. Ferroptosis inhibitors rescued cells from cell death.

Fig. 2

ARPE-19 cells were treated with DMOG and SI and co-treated with inhibitors of apoptosis, necroptosis, and ferroptosis. Treatment with A the apoptosis inhibitor Z-VAD increased DMOG + SI-induced cell death. B RIPK1 inhibitor Nec-1s, C RIPK3 inhibitor GSK872, and D MLKL inhibitor necrosulfonamide (NSA) are efficient inhibitors of necroptosis, but only RIPK3 inhibition by GSK872 fully rescued cells from cell death. Both ferroptosis inhibitors E Fer-1 and F Lip-1 fully rescued ARPE-19 cells from DMOG + SI-induced cell death. To further confirm ferroptotic cell death, cells were stained with the lipid peroxidation probe C11-BODIPY581/591 which revealed mainly perinuclear localization of lipid peroxidation-induced green fluorescence (G). H Quantification of cell fluorescence revealed significantly higher corrected total cell fluorescence in DMOG + SI treated cells compared to controls. Data are presented as means + SD. Inhibition assays were conducted with 6 independent groups (except for NSA treatment; N = 4) and statistical significance was calculated with two-way ANOVA. C11-BODIPY581/591 staining was assessed in 3 independent groups (n = 100) and statistical analysis was conducted with t-test. ns not significant, *p < 0.05, ***p < 0.001, and ****p < 0.0001. Scale bar = 50 μm.