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

Figure 6: DMT1(smf-3) KO protects against neuronal Aβ pathology:

Figure 6:

Neuronal Aβ worms exhibit increase sensitivity to iron toxicity than WT. A) Paralysis: Staged L4 worms (WT and neuronal Aβ) were transferred to plates containing iron (0 or 8.75 μM). 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). ns not significant, ****p<0.0001, one-way ANOVA, Tukey post hoc test. B) Non-paralyzed worm swimming rates. Staged L4 worms (WT and neuronal Aβ) were transferred to plate containing iron (0 or 8.75 μM). Worms were then transferred every 24 h for 5 days. 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. C) Neuronal Aβ iron sensitivity not mediated by iron burden. Tissue iron burden was measured in WT and Aβ treated with 8.75 μM iron for 5 days using ICP-MS. Data are mean ±SEM, N=3 independent biological replicates. ns not significant, Unpaired t test. D). Knock out of smf-3 in neuronal Aβ worms abolished paralysis. Staged L4 worms (neuronal Aβ and neuronal Aβ+ smf-3 KO) were transferred to plates containing iron (0 or 35 μM). 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, ****p<0.0001, one-way ANOVA, Tukey post hoc test. E) smf-3 KO in neuronal Aβ worms protects against Aβ decreased swimming rate. Staged L4 worms (neuronal Aβ and neuronal Aβ+ smf-3 KO) were transferred to plates containing iron (0 or 35 μM). Worms were then transferred every 24 h for 5 days. 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<0.001, ***p=0.0001, ****p<0.0001, one-way ANOVA, Tukey post hoc test.