Liver proteomes from WT, Rcrin knockout, AAV-Ctrl and AAV-Rpl8 mice

The liver tissues were disrupted by the ultrasonic processor on ice in lysis buffer. After centrifugation, the extracted proteins were reduced with 10 mM DTT for 2 h at room temperature followed by alkylation with 20 mM iodoacetamide for 30 min in the dark. The protein solution was diluted 1:5 with 50 mM triethylammonium bicarbonate (TEAB) and digested with trypsin (1:50) at 37 °C overnight. The digestion was desalted on OASIS HLB column and peptides eluted with 60% acetonitrile were lyophilized via vacuum centrifugation. All nanoLC-MS/MS experiments were performed on Orbitrap Eclipse (Thermo Scientific) equipped with an Easy n-LC 1200 HPLC system (Thermo Scientific). The DIA raw data from Orbitrap Eclipse were analyzed using Spectronaut version 17 (Biognosys) with the “DirectDIA” mode for protein identification and quantification. The Uniprot mouse protein database (updated on 03-2023) was used for searching the data from exosome samples.