Frozen postmortem human brain samples (BA40) were obtained via the National Institutes of Health (NIH) NeuroBioBank and stored at −80°C until use. Patient information provided by the NIH NeuroBioBank is listed in table S3. Frozen tissue samples were dissected to obtain approximately 50 mg of brain tissue, and tissue homogenates were generated as above for mouse brain tissues. Sarkosyl was added to each sample (500 μg of total proteins in 200 μl of total volume) for a 0.2% final concentration. The diluted protein samples were then incubated at 4°C for 30 min before ultracentrifugation at 100,000g for 1 hour at 4°C. Following ultracentrifugation, the supernatants were removed with the pellets resuspended and sonicated briefly in radioimmunoprecipitation assay (RIPA) buffer [50 mM tris (pH 7.4), 150 mM NaCl, 1% NP-40, 0.5% sodium deoxycholate, 0.1% SDS, and a protease inhibitor mix]. Proteins from total tissue homogenates and detergent-insoluble pellets were separated by SDS-PAGE as described above. All paired samples (paired by NIH NeuroBioBank for age, sex, and ethnicity) were loaded side by side for comparison in the order listed in table S3.

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