Quantitative proteomics reveals daf‐16‐mediated reduction in protein metabolism in long‐lived daf‐2(e1370) mutants. Quantitative proteomics reveals daf‐16‐mediated reduction in protein metabolism in long‐lived daf‐2(e1370) mutants. (A) Schematic overview of experimental set‐up. Protein extracts were generated from three sample sets as biological replicates, harvested at day 1 of adulthood. Proteins were trypsin digested and labelled with unique mass tags. Labelled peptides were combined, fractionated by strong cation exchange chromatography to reduce sample complexity and subsequently analysed by LC‐MS/MS. Data from 66 fractions were merged and analysed using Proteome Discoverer software. (B) Percentage of proteins with differential abundance (>1.3 fold change) in daf‐2(e1370) mutants compared with N2 (left bar), and their expression dependency on daf‐16 (right bars) for proteins with increased and decreased abundance to N2, respectively. The number of quantified proteins out of total amount of identified proteins is indicated on top of the bars. (C) Boxplots show expression analysis for GO‐enriched processes in daf‐2(e1370) and daf‐16(mu86); daf‐2(e1370) mutants relative to N2 (Log2). Asterisks indicate statistical significance (*P<0.01, **P<0.001, ***P<0.0001) as determined by paired t‐test analysis. Gerdine J Stout et al. Mol Syst Biol 2013;9:679 © as stated in the article, figure or figure legend