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MINNEAPOLIS, MN - FEBRUARY 04: Nick Foles #9 of the Philadelphia Eagles celebrates a 21-yard touchdown during the second quarter against the New England Patriots in Super Bowl LII at U.S. Bank Stadium on February 4, 2018 in Minneapolis, Minnesota. (Photo by Elsa/Getty Images)

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https://doi.org/10.1371/journal.pbio.1000450.g004

Overall, our data show that age-dependent protein aggregation is not necessarily caused by increased protein levels. Instead, other events associated with age can promote aggregation.

Protein Insolubility in Young Animals

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To further evaluate the importance of the aging process in mediating protein aggregation, we asked whether a mutation that slows aging would affect the rate of inherent protein aggregation. Mutation of the daf-2 /insulin/IGF-1-receptor gene doubles the lifespan of C. elegans Cashmere Silk Scarf amity by VIDA VIDA tCUosrrL
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. The pattern of insoluble proteins in young-adult animals subjected to daf-2 RNAi (from the first larval stage) resembled that of wild type ( Figure 5A ). However, we found only a slight increase in insolubility with age (1.6-fold) compared to control animals (3.6-fold). More strikingly, the insolubility pattern of proteins extracted from daf-2(RNAi) animals did not change with age, whereas the two patterns (young versus old) were very different in wild type. Exposure to daf-2 RNAi from the last larval stage produced very similar results (unpublished data). Similarly, Western blot analysis showed that daf-2 RNAi treatment greatly reduced the extent of both RHO-1 and DAF-21 insolubility with age and moderately reduced the extent of KIN-19 and PAR-5 age-dependent insolubility ( Modal Scarf SweetCatheads by VIDA VIDA Wo9n7C8Y9m
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