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. 2019 May 21;10:2191. doi: 10.1038/s41467-019-10136-w

Fig. 1.

Fig. 1

Long-lived proteome analysis during Drosophila aging. a Experimental scheme of metabolic labeling in Drosophila. Flies were first fully labeled with 15N, and then switched to normal 14N food at 5d. Samples were collected at 5d, 30d, and 60d of age. Proteomes of three tissues with indicated ages, and the ubiquitylome of 60d old head and muscle tissues were analyzed. The proportion of 15N signal represents protein turnover rate. b 15N proportions (15N %) for each protein in different tissues and different aging time point. Proteins quantified during aging were compared across tissue types. A high 15N % value at aging time point represents a slow turnover rate. The protein identifiers were used for comparison among multiple datasets in this study. c Clustering of proteins according to their 15N proportion changes during aging. Proteins in cluster 1 were characterized with the slowest turnover rates thus defined as extremely long-lived proteins (ELLPs). The median values are marked at the center. The boxes extend from the 25th to 75th percentiles. The upper or lower whiskers extend from the hinge to the largest or smallest value no further than 1.5 times the interquartile range (IQR).d Venn diagram and GO analysis of ELLPs in head and muscle. 149 ELLPs were found in both head and muscle, two post-mitotic tissues, and then subjected to functional analysis, with “nucleosome assembly” as the most significant hit (p= 5.2e-9, Fisher’s exact test). e H2A and H2B are ELLPs. H2A and H2B belonging to “nucleosome assembly” category were identified with a dramatic enrichment of 15N signals at day 60. The extracted ion chromatographs (XICs) of the paired 15N and 14N peaks from two representative peptides were compared