Endogenous complexes for LC-MS/MS were affinity-purified following the IP protocol. Quality of the samples was determined by silver staining and Western blot. Briefly, the SDS-PAGE gel was fixed in fixation buffer 1 (50% methanol and 10% acetic acid) for at least 15 min and fixation buffer 2 (10% methanol and 7% acetic acid) for 2 hours. The gel was washed with gluteraldehyde (25%):water solution (1:10) for 15 min and then three times with deionized water for 15 min. The staining solution was prepared by dropping solution B (1 g of AgNO3 in 5 ml of deionized water) into solution A (0.185 ml of 10 M NaOH, 2.8 ml of NH4OH, and 22.5 ml of deionized water). The final volume was brought to 100 ml by adding 70 ml of deionized water. The gel was stained for 15 min and washed three times with deionized water for 2 min. The stain was developed with developing solution (0.5 ml of 1% citric acid and 0.05 ml of 38% formaldehyde in 100 ml of deionized water) to appropriate signal and then stopped by stop solution (50% methanol and 5% acetic acid) for 10 min.

LC-MS/MS was performed by The Wistar Proteomic Facility. MS/MS spectra generated from the LC-MS/MS runs were searched using full tryptic specificity against the UniProt human database using the MaxQuant 1.5.2.8 program. Protein quantification was performed using unique and razor peptides. Razor peptides are shared (nonunique) peptides assigned to the protein group with the most other peptides (Occam's razor principle). False discovery rates (FDR) for protein and peptide identifications were set at 1%.

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