Evi1 interacts with PcG proteins. (A-C) Immunoprecipitation of HA-Evi1 identified EZH2 (A), SUZ12 (B), and EED (C) as interacting proteins in 293T cells. Vertical lines have been inserted to indicate a repositioned gel lane in panel A. (D) Interaction between Evi1 and endogenous PRC2/3/4 proteins in Evi1-induced murine leukemic cells. (E) Interactions between endogenous Evi1 and PRC2/3/4 in human leukemia cells derived from the patient with AML patient. (F-I) Immunoprecipitation of HA-Evi1 identified BMI1 (F), RING1 (G), RING2 (H), and HPH2 (I) as interacting proteins in 293T cells. A vertical line has been inserted to indicate a repositioned gel lane in panel I. (J) ZF1-7 domain of Evi1 is responsible for the physical interaction with EZH2. IgG indicates immunoglobulin G; WB, Western blotting.