Fig. 7. nduf-2.1 Cys366 effects on hypoxic signaling.

a Complex I ROS requires Cys366 for acute hypoxic signaling. Staged L4 wild-type, nduf-2.1(C366S), nduf-2.1(C366D) worms were singled on to an unseeded plate and baseline body bends were counted. Worms were then subjected to a short period of hypoxia and subsequent reoxygenation. Data are mean ± SD. N = 15 independent animals across 3 technical replicates, *p < 0.05, **p < 0.01 (Two-way RM ANOVA, Dunnett’s multiple comparisons). b Complex I ROS requires Cys366 for hypoxic signaling and protection against hypoxia reperfusion injury. Staged L4 wild-type, nuo-1::Supernova, nduf-2.1(C366S), nduf-2.1 (C366D) worms were subjected to hypoxia reperfusion injury and survival was assessed. Where indicated, worms were exposed to a hypoxic preconditioning stimulus. Data are mean ± SD. N = 4–19 independent replicates each containing 25–100 animals **p < 0.01 (two-way ANOVA, Sidak’s multiple comparisons). c Mean survival difference was calculated from data in (b). Mean survival difference was calculated by subtracting the survival from hypoxia reoxygenation group from the preconditioning group for each daily technical replicate, Data are mean ± SD. N = 4–19 independent replicates, *p < 0.05, **p < 0.01 (one-way ANOVA, Dunnett multiple comparisons). d Schematic illustration of different light treatments pre- and post-hypoxic conditions. e Complex I ROS is sufficient to protect against hypoxic injury. Stage L4 wild-type, nuo-1::Supernova worms were exposed to light (Quantum, 0.02 mW/mm²) as indicated in (d). Light was given for 4 h during the preconditioning period (PC-period), 4 h immediately prior to the hypoxic insult, 21 h immediately prior to the hypoxic insult, or 4 h immediate after the hypoxic insult. Survival was scored 24 h post-reoxygenation. Data are mean ± SD. N = 9–23 independent replicates each containing 25–100 animals, *p < 0.05, **p < 0.01 (Two-way ANOVA, Sidak’s multiple comparisons).