Figure 2.

AKT-mediated EZH2 phosphorylation at S21 induces EZH2 to interact with RNAPII

(A) Western blots of EZH2 immunoprecipitations (IP) performed using nuclear lysates prepared from SW480 cells untreated or treated with 250 μM H₂O₂ for 30 min. IP with IgG serves as a negative control. Input is nuclear lysates used for IP.

(B) EZH2 IP performed using nuclear lysates prepared from SW480 cells treated with 10 μM AKT inhibitor (AKTi, GSK-690693) or DMSO for 48 h followed by no additional treatment or co-treatment with 250 μM H₂O₂ for 30 min. IP with beads only serves as a negative control. See Figure S2A for western blots of whole-cell extract (WCE) and input.

(C) AKT IP performed using nuclear lysates prepared from EZH2 knockdown (KD) and empty vector (EV) SW480 cells treated as in A.

(D) HA IP performed using nuclear lysates prepared from HA-EZH2 wildtype (WT) and HA-EZH2 phospho-null (PN) S21A expressing SW480 cells treated as in A. IP with IgG serves as a negative control. See also Figure S2.