293T NME4 TAP-MS and TurboID-MS

To identify interacting proteins of NME4 in human cell lines, we using tandem affinity purification followed by mass spectrometry (TAP-MS) in HEK293T cells. We established stable cell line which expressing NME4 gene fused with SFB triple tags (S tag-2x Flag tag-SBP tag) or TurboID. We isolated, combined, and affinity-purified the membrane and soluble fractions using TAP. We identified proteins associated with the bait in the isolated complexes using MS and searched the uniprot human database for these proteins.