Figure 1.

AKT activation mediates EZH2 phosphorylation at Serine 21

(A) Western blots of AKT immunoprecipitations (IP) performed using nuclear lysates prepared from EZH2 knockdown (KD) and empty vector (EV) SW480 cells untreated or treated with 250 μM H₂O₂ for 30 min. See Figure S1A for western blots of nuclear lysates used for IP (input) and total protein prior to cellular fractionation (whole-cell extract-WCE).

(B) EZH2 IP performed using nuclear lysates prepared from SW480 cells untreated or treated with 10 μM AKT inhibitor (AKTi, GSK-690693) for 48 h followed by no additional treatment or co-treatment with 250 μM H₂O₂ for 30 min.

(C) 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.

(D) AKT IP performed using nuclear lysates prepared from empty vector (EV) and PTEN knockdown (KD) Caco2 cells.

(E) AKT IP performed using nuclear lysates prepared from EV and PTEN KD RKO cells. See also Figure S1.