ESCO2 Protein Complex Identification

HEK293T cells were transfected with Flag-ESCO2 vector for 48 h using Lipofectamine 2000. Treated HEK293T cells were lysed in Co-IP lysis buffer (P0013, Beyotime, Shanghai, China). Co-IP was performed using anti-FLAG/anti-HAantibodiesand protein A/G agarose beads (sc-2003, Santa Cruz) to extractthe complexes. Gel bands were detected using a silver staining kit (P0017S, Beyotime) combined with MS following the manufacturer’s protocol. According to a previously published method [25 (link)], the peptides of the bands were analyzed using nano-LC–MS/MS. The mass spectrometry proteomics data have been deposited to the ProteomeXchange Consortium via the PRIDE partner repository with the dataset identifier PXD023527 and PXD23600.