Nanolc ms ms

Co-immunoprecipitated proteins were visualized using a Coomassie Blue Staining Kit (Beyotime, China). The protein bands of interest were meticulously excised from the gel and subsequently digested. These peptides were analyzed with nanoLC-MS/MS (Sangon Biotech, Shanghai, China), using an Orbitrap Fusion mass spectrometer. The mass spectrometry-generated raw spectral data were then processed and analyzed employing PEAKS Studio 8.5 software for detailed characterization.

Co-IP proteins were visualized by Fast Silver Stain Kit (Beyotime, China). The protein bands of interest were excised from the gel and subjected to in-gel trypsin digestion. Resulting peptides were extracted and analyzed by nanoLC-MS/MS (Sangon Biotech, Shanghai, China) using an Orbitrap Fusion mass spectrometer. The shotgun strategy was employed for proteomics studying. Protein identification was performed using Mascot search engine and the SwissProt database. The mass spectrometry proteomics data have been deposited to the ProteomeXchange Consortium (http://proteomecentral.proteomexchange.org) via the iProX partner repository [39 (link), 40 (link)] with the dataset identifier PXD041447.