Ribonucleic acid from

Unlabeled standards were prepared for all relevant analytes, and were added directly to the biomass pellet prior to the execution of the protocols described below. For amino acids, 40 µL of a 2.5 mM per amino acid solution (Pierce 20088) was used. For RNA, a 1 mg/mL solution (ribonucleic acid from torula yeast, Sigma R6625) in water was prepared, of which 80 µL was added to the sample. For glycogen, a 0.1 mg/mL solution (glycogen from bovine liver, Sigma G0885) in water was prepared, of which 100 µL was added to the sample. For fatty acids, a solution of 0.3 mg/mL of myristic acid (C14:0), palmitic acid (C16:0), palmitoleic acid (C16:1), stearic acid (C18:0), and elaidic acid (C18:1) in hexane was prepared, of which 20 µL was added.

In order to maintain strains and collect flies for lifespan studies, we used corn meal food (85.7 g corn meal “Aunt Jemina” (The Quaker Oats Company, Chicago, IL), 50 ml golden A unsulfured molasses (Groeb Farms Inc, Onsted, MI), 71.4 g Torula yeast (MP Biomedicals, Solon, OH), 2.86 g p-hydroxybenzoic acid methyl ester (Sigma), 6.4 g agar (MoorAgar Inc, Loomis, CA) and 5.7 ml propionic acid (Sigma) per liter water). In order to test lifespan of flies, fully defined medium was used based on the fruit fly basal mix developed in a recent study [19 (link)] with modification in amino acid content. This diet contained 62.08 g Diet TD.04310 (corresponds to the 0.1X diet) and/or 101.07g Diet TD.10417 (corresponds to the 1.0X diet) (Harlan Teklad), 100 mg lecitin from soybean (Sigma), 500 mg ribonucleic acid from Torula yeast (Sigma), 100 g of dextrose, 20 g agar, 2.85 ml propionic acid, 0.255 ml of phosphoric acid (Sigma), and indicated amounts of Met (Sigma) per liter of water. To prepare 0.4X and 0.7X diets, the two basal mixes were mixed to adjust amino acid concentrations.