Fig. 3.

Both As³⁺ and H₂O₂ induce the interaction of Akt and EZH2 and Akt-dependent phosphorylation of the exogenous EZH2 overexpressed in HEK-293 cells. (A) HEK-293 cells were transfected with 3myc-tagged EZH2 expression vector for 24 h followed by the treatment of the cells with 20 µM As³⁺ for 2 h in the presence or absence of 10 mM NAC, followed by Western blotting using the indicated antibodies. (B) The transfected HEK-293 cells were treated with As³⁺ or H₂O₂, followed by Western blotting as in (A). (C) Anti-myc tag or control IgG was used in immunoprecipitation (IP) to pull down the exogenousmyc-tagged EZH2. The IP was then subjected to Western blotting using antibodies recognizing the phosphorylated Akt substrate motif (RXRXXS*/T*), myc tag, pAkt, and Akt.