Trigonelline chloride

WT hermaphrodite Bristol worms (N2) and GFP-tagged myosin worms^{81 (link)} were treated with trigonelline chloride (catalogue no. T5509, Sigma-Aldrich) and NR chloride from day 1 of adulthood. Nematodes were cultured at 20 °C on nematode growth medium (NGM) agar plates seeded with Escherichia coli strain OP50. Trigonelline and the other precursors were added to the NGM medium at a final concentration of 1 mM just before pouring the plates, unless otherwise stated. An enzymatic method adapted from Dall et al.^{71 (link)} was used to measure NAD⁺ content in worms. Fifty to 100 worms were collected per biological replicate and NAD⁺ levels were normalized on the protein content. The bacterial feeding RNAi experiments were carried out as described^{82 (link)}. RNAi was performed using the clones sir-2.1 (R11A8.4) and nprt-1 (Y54G2A.17) from Geneservice.

All solvents used were of HPLC grade and supplied by Fisher Scientific. Canavanine sulfate salt, quercetin-3-glucoside, quercetin dihydrate, hyperoside, luteoloside, stachydrine (hydrochloride), trigonelline chloride, diosmetin, H-Glu-Tyr-OH and H-Phe-Glu-OH standards were purchased from the Sigma-Aldrich (Munich, Germany), Thermo scientific, Carl Roth (Karlsruhe, Germany), Chemodex (St. Gallen, Switzerland), Cayman (Düsseldorf, Germany) and Adipogen life sciences (San Diego, CA, USA) companies.