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. 2022 Apr 21;4(3):fcac103. doi: 10.1093/braincomms/fcac103

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© The Author(s) 2022. Published by Oxford University Press on behalf of the Guarantors of Brain.

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.

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Differential centrifugation can be used to separate proteins into consistent fractions in post-mortem human brain. (A) Schematic of the centrifugation scheme used to prepare samples for this experiment. Centrifuge speeds and spin times are provided. All spins were performed at 4°C. (B) PCA shows good separation of samples by centrifugation fraction in the first two principal components. (C) An Upset plot shows that most proteins in the data set differentially expressed by ANOVA are differentially abundant between Fraction 7 (cytosolic fraction) and Fraction 6 (large protein complex fraction) and all other fractions. (D) A heatmap of differentially expressed proteins (ANOVA, see Supplementary Table 3 for test statistics) shows that samples generally cluster on the basis of centrifugation fraction. The exception is Fractions 2 and 3, where samples from the same individual cluster in pairs within the larger cluster. Colour coding of samples in the horizontal bar is identical to colour coding in B.