2 deoxy 5 fluorouridine fudr

After age synchronization, we transferred 40 young adult nematodes to fresh NGM plates. In addition, 2′-deoxy-5-fluorouridine (FUDR) (Sigma, St. Louis, MO, USA) was added to the plate to block C. elegans reproduction. QBLP was added to liquid NGM at three different final concentrations: 0.02, 0.2 and 2 g/L. Lifespan is monitored daily from the first day of adulthood. The numbers of live and dead worms were recorded every two days until all the worms were dead. The animals were considered to be dead when no pharyngeal pumping was observed or when the nematodes did not respond to touch. The mean and median lifespans were calculated. Three replicates were analyzed per exposure group [26 (link),27 (link)].

Neuron cultures were obtained from C57BL/6 male mice of 8 weeks. All experimental procedures were carried out in compliance with the Animal Research: Reporting of In Vivo Experiments (ARRIVE) guidelines and in accordance with National Institute of Health guidelines for animal care and use of Laboratory animals (DL 2016, Italian Ministry of Health approval protocol 919/2015-PR). A complete dorsal root ganglia (DRG) pool was collected in F12 medium (Euroclone, Italy) from each mouse. DRG were digested for 1 h with 12.5 mg/ml collagenase (Sigma Aldrich, USA) and 10 mg/ml DNase (Sigma Aldrich, USA) and then mechanically triturated. A BSA gradient (30% BSA, 70% F12 medium) was used to isolate neurons that were then resuspended in BS medium (F12 medium supplemented with 1% N2 supplement 100X (Life Technologies, UK), 1% BSA (Sigma Aldrich, USA), 1% Penicillin and Streptomycin 100X (Euroclone, Italy), 1% L-glutamine 100X (Euroclone, Italy) and seeded in a single drop on poly-l-lysine (Sigma Aldrich, USA) coated dishes. After 24 h, neurons cultures were treated with 10⁻⁵ M 2′-Deoxy-5-fluorouridine (FuDR) (Sigma, USA) to remove satellite cells.