Figure 3
From: Protein Phosphatase 2A (PP2A) Regulates EG5 to Control Mitotic Progression
PP2A dephosphorylates EG5 at Threonine 926. (A) Intensity of peptide containing the phosphorylated Thr 926 site in PP2A/C or control protein treated EG5. (B) PP2A/C dephosphorylated EG5 in a concentration-dependent manner. Indicated amounts of His-PP2A/Cα were titrated into a constant amount of FLAG-EG5, reacted in phosphatase buffer at 30 °C for 30 minutes, and the mixture was precipitated using anti-FLAG antibody and subjected to Western blot. (C) Phosphorylation levels of EG5 (T926) in PP2A/Cα knockdown and control cells. (D) Protein levels of PP2A/C and phosphorylation level of EG5 (T926D) in HeLa cells with or without NOC treatment. rPP2A/Cα represents shPP2A/Cα resistant PP2A/Cα. (E) Phosphorylation levels of EG5 (T926) in PP2A/B55α knockdown and control cells. (F) Phosphorylation levels of EG5 (T926) and protein levels of PP2A/C or PP2A/B55α in HeLa cells overexpressing PP2AC/α or PP2A/B55α respectively. For the original uncropped western blot data, see Supplementary information.
