Phosphorylation of EZH2 increases its binding to chromatin
(A) Western blots of chromatin lysate prepared from HA-EZH2 wildtype (WT), HA-EZH2 phospho-null (PN) S21A and HA-empty vector (EV) expressing SW480 cells treated with 250 μM H2O2 for 30 min. Whole-cell extract (WCE) serves as a control for the chromatin extract.
(B) Western blots of chromatin lysate prepared from HA-EZH2 WT, HA-EZH2 S21D and HA-EV expressing SW480 cells.
(C) Western blots of chromatin lysate prepared from SW480 cells untreated and treated with 1 μM EZH2 inhibitor (EZH2i, GSK-503) for 72 h or with 10 μM AKT inhibitor (AKTi, GSK-690693) for 48 h and followed by no additional treatment or co-treatment with 250 μM H2O2 for 30 min.
(D) Western blots of chromatin lysate prepared from FLAG-β-catenin WT and FLAG-β-catenin K49R expressing PTEN knockdown (KD) and EV expressing SW480 cells. WCE serves as a control for chromatin extract.