MSLN has only been implicated to have one binding partner, MUC16/CA125. In an effort to identifying novel binding partners of MSLN, an IP-MS experiment was conducted using immunoprecipitations of MSLN in whole cell lysate of NOMO-1 and MSLN KO cells. We identified Src-family kinase member Lyn, and guanine nucleotide-binding protein G(i), alpha subunit proteins, GNAI1, GNAI2, and GNAI3 as a novel binding partners of MSLN in AML.