Phosphoproteomics of Hela cells stimulated with EGF in the presence of protein phosphatase inhibitors

We profiles the effects of six protein phosphatase inhibitors on EGF-dependent phosphoproteome dynamics. The cells were treated with 20 nM EGF for 8 or 20 min with or without 15 min pre-treatment of one of the phosphatase inhibitors (100 nM or 1 M; Raphin1 acetate, sanguinarine chloride, KY-226, SHP099, NSC95397, or BCI). We further focused on sanguinarine chloride, and Hela cells were treated with 20 nM EGF for 3, 8, or 20min with or without 15 min pre-treatment of sanguinarine (5, 50, or 500 nM). We also examined combination of sanguinarine chloride (500 nM) and BIRB796 (500 nM), a p38-kinase inhibitor.