Fig 3. A diverse group of short-sleeping mutants is sensitive to oxidative stress.

We asked (A) whether other sleep mutants unrelated to inc or Cul3 share the same sensitivity to oxidative stress. (B–D, left panels) We found that sleepless mutants slept 65% less than controls (B, p < 0.0001, n = 6–10 flies/genotype), fumin mutants slept 95% less than controls (C, p < 0.0001, n = 15–16 flies/genotype), and redeye mutants slept 50% less than controls (D, p < 0.0001, n = 16 flies/genotype). (B–D, middle panels) When injected with paraquat, sleepless mutants (B, p < 0.0001, n = 100 flies/genotype), fumin mutants (C, p < 0.0001, n = 97–98 flies/genotype), and redeye mutants (D, p < 0.0001, n = 88–92 flies/genotype) died faster than controls. (B–D, right panels) Faster death kinetics were also observed after H₂O₂ feeding relative to controls for sleepless mutants (B, p < 0.0001, n = 40 flies/genotype), fumin mutants (C, p < 0.0001, n = 39–40 flies/genotype), and redeye mutants (D, p < 0.0001, n = 39–42 flies/genotype). For scatterplots (B–D), each data point represents the average sleep in minutes/day measured across 4–5 days for an individual animal. Data are shown as mean ± SEM and p-values were obtained by ordinary one-way ANOVA followed by a post hoc Tukey test when significance was detected. For survival curves (B–D), p-values were obtained by log-rank analysis. Data from representative experiments are shown. Each experiment was performed at least three times. Raw data from representative experiments are available in S1 Data; raw data from all trials are available upon request. Cul3, Cullin-3; inc, insomniac.