*2.3. Stress Assays*

Oxidative stress in worms was induced by subjecting the animals to 35 ◦C heat-shock treatment. Worms were incubated at 20 ◦C on NGM-*E. coli* OP50 plates with or without Q until days 2 and 9 of adulthood. Then they were transferred with a platinum wire to agar plates Ø 35 mm, 20 worms per plate) and switched to 35 ◦C for 4, 6, or 8 h. The time was decided depending on the thermotolerance of the specific worm strain used in the assay, which was previously checked. After that time, dead and alive nematodes were counted. In the studies involving the use of worm mutants, in addition to the mutant control, a parallel control using N2 wild-type (WT) worms was also included. For the assays, ten plates were used per treatment containing 20 worms per plate, resulting in a total of 200 worms, although a small percentage of worms was usually lost in the score. Only in the case of the *skn-1* mutant (EU1) were 300 worms (20 worms per plate/15 plates) used. According to the information supplied by the CGC, the work with this mutant required special considerations, as only the homozygote worms with WT appearance are considered for the assay, while the uncoordinated heterozygote worms are only employed to maintain the lineage. To have reproducible results in the heat shock assays, that is, to have the smallest possible temperature oscillations, some precautions were followed, in agreement with the guidelines stated in the reference paper [25]. During assays, the door of the incubator was only opened when the survival rate had to be measured. The plates were not stacked inside the incubator, they were placed in a row on the same shelf of the incubator, leaving enough space between them for the air to circulate properly. The temperature was controlled with the incubator's own thermostat and with an external thermostat with sensors inside the incubator to monitor possible temperature oscillations.
