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[Preprint]. 2024 Aug 9:2024.08.08.607074. [Version 1] doi: 10.1101/2024.08.08.607074

Figure 4: Ferroptosis modulates iron induced toxicity.

Figure 4:

A). Schematic diagram showing side of ferroptotic drug target regulating iron toxicity. B). Ferrostatin-1 (Fer-1) ameliorates iron-induced worm paralysis. Staged L4 worms were transferred to plates containing 0 μM iron, 0 μM iron + 5 μM Fer-1, 35 μM iron and 35 μM iron + 5 μM Fer-1. Paralysis was scored every 24 h for 5 days. Data are mean ±SEM, N=5 independent biological replicates (where one biological replicate contains 20 worms per plate). ****p<0.0001, one-way ANOVA, Tukey post hoc test. C). Ferrostatin-1 restored non-paralyzed worm swimming rates in iron toxicity environment. Staged L4 worms were transferred to plates containing 0 μM iron, 0 μM iron + 5 μM Fer-1, 35 μM iron and 35 μM iron + 5 μM Fer-1. Non-paralyzed worms were individually transferred to plate containing 100 μl of buffer. After 30 seconds of equilibration, swimming rates were collected for 15 seconds. Data are mean ±SEM N=5 independent replicates (where 4 independent worm count constitute an N). ns not significant, ****p<0.0001, one-way ANOVA, Tukey post hoc test. D). RSL3 exacerbates iron toxicity induced increase in worm paralysis. Staged L4 worms were transferred to plates containing 0 μM iron, 0 μM iron + 5 nM RSL3, 0 μM iron + 5 nM RSL3 + 5 μM Fer-1, 35 μM iron, 35 μM iron + 5 nM RSL3, and 35 μM iron + 5 nM RSL3 + 5 μM Fer-1. Paralysis was scored every 24 h for 5 days. Data are mean ±SEM, N=5 independent biological replicates (where one biological replicate contains 20 worms per plate). ****p<0.0001, one-way ANOVA, Tukey post hoc test. E). RSL3 worsen non-paralyzed worm swimming rates in iron toxicity environment. Staged L4 worms were transferred to plates containing 0 μM iron, 0 μM iron + 5 nM RSL3, 0 μM iron + 5 nM RSL3 + 5 μM Fer-1, 35 μM iron, 35 μM iron + 5 nM RSL3, and 35 μM iron + 5 nM RSL3 + 5 μM Fer-1. Non-paralyzed worms were individually transferred to plate containing 100 μl of buffer. After 30 seconds of equilibration, swimming rates were collected for 15 seconds. Data are mean ±SEM N=5 independent replicates (where 4 independent worm count constitute an N). ****p<0.0001, one-way ANOVA, Tukey post hoc test