Figure 5

HyPer ratios in WT animals and nox mutants

WT animals and nox mutants were generated in a Tg(ubb:HyPer)umc400 transgenic background, which permits in vivo assessment of ROS levels. (A) ROS Hyper ratios in individual WT animals (n = 8) observed at three time points during the day over three days post-amputation reveal an oscillatory pattern of ROS levels.

(B) ROS levels over three consecutive days post-amputation were significantly higher in the mornings (7a.m.) than in the afternoons (3 p.m.) or nights (10p.m.).

(C and D) Long-Scargle periodogram reveals that the ROS oscillations follow a daytime-dependent trend, with a periodicity of around 0.4 (∗ on graph), indicative of a ∼24 h circadian cycle (D).

(E) Graph of ROS levels measured at the same time of day (3p.m.) in WT (unamputated), WT (amputated), cyba^umc403, duox^sa9892, and nox5^umc402 animals.

(F) Graph of mean Hyper ratios measured over three consecutive days at 3p.m. reveals significantly attenuated post-amputation ROS levels among the duox^sa9892 mutants relative to the WT amputated fins and the other nox mutants. Indeed, ROS levels in the duox^sa9892 fins are significantly lower than even those found in unamputated WT fins. ROS levels in the amputated cyba mutant fins are significantly lower than those present in WT amputated fins but not as low as in the duox^sa9892 fins. ROS levels in the amputated nox5^umc402 mutant fins are not significantly different from those present in amputated WT fins. (Bonferroni’s multiple comparisons test, ∗p < 0.5, ∗∗p < 0.01, ∗∗∗∗p < 0.0001). For A-D, WT (n = 8). For E-F, WT unamputated (n = 7); WT amputated (n = 8); cyba^umc403 (n = 10); duox^sa9892 (n = 7); nox5^umc402 (n = 7).

See also Figures S10 and S11.