*2.4. RT-qPCR Assays*

Adult worms of the N2 *C. elegans* strain were treated with or without 200 μM of Q for 4 days. The worms were collected with M9 buffer, centrifuged at 10,000 *g* for 1 min, and the pellet was dissolved in 300 μL of M9, to which 3.5 μL of 2-mercaptoethanol was added. Total RNA was extracted using the RNAspin Mini RNA Isolation Kit (GE Healthcare). In order to maximize cell breakage, in the first stage of the extraction, 10 stainless-steel beads (2 mm) were added. The mixture was vortex shaken vigorously and further homogenized in a Thermo Savant FastPrep 120 Cell Disrupter System, with a speed of 5.5 m/s and run time duration of 10 s, five times. cDNA was produced with high-capacity cDNA reverse transcription kit (Applied Biosystems) using 2 μg of total RNA per reaction. The expression of mRNA was assessed by quantitative real-time PCR, using SYBR green as

the detection method. The gene expression data were analyzed using the comparative 2−ΔΔCt method, with *act-1* as the normalizer [26]. Nine independent experiments were performed. *Act-1* was used as a normalizer both in the assays carried out in non-stressed and stressed worms. The information related to gene-specific primers used in this work can be found in the Supplementary Table S1.
