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. 2020 Sep 29;10:15983. doi: 10.1038/s41598-020-72410-y

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© The Author(s) 2020

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

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(A) Representative immunoblot of fractionated brain tissue. A = unfractionated brain, B = nuclear fraction, C = synaptosomal fraction, and D = mitochondrial fraction. Fractionation was validated by the enrichment of proteins known to reside in each compartment: HDAC2(nuclear protein), SynGAP(synaptic protein), and COX IV (mitochondrial protein). The images represent five separately processed immunoblots. The uncropped immunoblot images are in Fig. S9. (B) Quantitation of the CCT5 and 8 IR in the synaptosomal (B) and nuclear (C) fraction normalized to the CCT IR in unfractionated brain; N = 4. (D) The number of AHA peptides identified from a 7 day chase period was decreased in 2-month old brains compared with 1-year old brain. The average percentages of heavy AHA peptide identified (y-axis) from the total AHA identifications (i.e. light plus heavy) were calculated from MS analysis. N = 3. One-ANOVA analysis with Bonferroni’s post-hoc test was performed. Figure depicts the Bonferroni p-values for 1-year old brain. *p < 0.05, **p < 0.01, ***p < 0.001,****p < 0.0001. Bonferroni p-values not in the figure: 2mo-Brain vs. 2mo-Liver **, 2mo-Brain vs. 1 yr-Liver **, and 2mo-Liver vs. 1 yr-Liver not significant. (E) AHA proteins have a slower degradation rate in brains from 1-year old mice than 2-month old mice. (F) AHA proteins have similar degradation rate in livers from 1-year old mice than 2-month old mice. x-axis is the log₂ fold change plotted as 1-year/2-month. Each point represents the average Day7/Day0 protein ratio, calculated from three mice in each age group. The y-axis is the log₁₀ p-value with the red line representing the significant value filter 0.05.